6-substituted 2-(benzimidazolyl)purine and purinone derivatives for immunosuppression

ABSTRACT

The present invention provides novel purinones and purines useful for the prevention and treatment of autoimmune diseases, inflammatory disease, mast cell mediated disease and transplant rejection. The compounds are of the general formulae I and II shown below, in which Q is selected from the group consisting of CX and nitrogen; and A is chosen from the group consisting of H, (C 1 -C 6 ) alkyl, heteroaryl, and aryl:

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority from U.S. Provisional Application Ser.No. 60/828,165, filed Oct. 4, 2006, and U.S. Provisional ApplicationSer. No. 60/828,169 filed Oct. 4, 2006, the entire contents of both areincorporated herein by reference.

FIELD OF THE INVENTION

The invention relates to 6-substituted 2-(benzimidazolyl)purine orpurinone derivatives and 6-substituted2-(imidazolo[4,5-c]pyridinyl)purine or purinone derivatives useful asimmunosuppressants.

BACKGROUND OF THE INVENTION

Immunosuppression is an important clinical approach in treatingautoimmune disease and in preventing organ and tissue rejection. Theclinically available immunosuppressants, including azathioprine,cyclosporine and tacrolimus, although effective, often cause undesirableside effects including nephrotoxicity, hypertension, gastrointestinaldisturbances and gum inflammation. Inhibitors of the tyrosine kinaseJak3 are known to be useful as immunosuppressants (see U.S. Pat. No.6,313,129).

The members of the Janus kinase (Jak) family of non-receptorintracellular tyrosine kinases are components of cytokine signaltransduction. Four family members have been identified to date: Jak1,Jak2, Jak3, and Tyk2. The Jaks play a key role in the intracellularsignaling mediated through cytokine receptors. Upon binding of cytokinesto their receptors, Jaks are activated and phosphorylate the receptors,creating docking sites for other signaling molecules, in particularmembers of the signal transducer and activator of transcription (STAT)family. While expression of Jak1, Jak2 and Tyk2 is relativelyubiquitous, Jak3 expression is temporally and spatially regulated. Jak3is predominantly expressed in cells of hematopoietic lineage; it isconstitutively expressed in natural killer (NK) cells and thymocytes andis inducible in T cells, B cells and myeloid cells (reviewed in Ortmann,et al., 1999 and Yamaoka, et al., 2004). Jak3 is also is expressed inmast cells, and its enzymatic activity is enhanced by IgE receptor/FcεRIcross-linking (Malaviya and Uckun, 1999).

A specific, orally active Jak3 inhibitor, CP-690,550, has been shown toact as an effective immunosuppressant and prolong animal survival in amurine model of heart transplantation and a primate model of kidneytransplantation (Changelian, et al., 2003). Furthermore, aberrant Jak3activity has been linked to a leukemic form of cutaneous T-cell lymphoma(Sezary's syndrome) and acute lymphoblastic leukemia (ALL), the mostcommon form of childhood cancer. The identification of Jak3 inhibitorshas provided the basis for new clinical approaches in treating leukemiasand lymphomas (reviewed in Uckun, et al, 2005). Two dimethoxyquinazolinederivatives, WHI-P131 (JANEX-1) and WHI-P154 (JANEX-2), have beenreported to be selective inhibitors of Jak3 in leukemia cells (Sudbecket al., 1999).

Jak3 has also been shown to play a role in mast-cell mediated allergicreactions and inflammatory diseases and serves as a target inindications such as asthma and anaphylaxis.

Therefore, compounds that inhibit Jak3 are useful for indications suchas leukemias and lymphomas, organ and bone marrow transplant rejection,mast cell-mediated allergic reactions and inflammatory diseases anddisorders.

SUMMARY OF THE INVENTION

It has now been found that compounds of general formula I and II arepotent and selective inhibitors of Jak3:

In these compounds,

-   Q is selected from the group consisting of CX and nitrogen;-   X is selected from the group consisting of hydrogen, halogen, and    electron-withdrawing groups;-   A is chosen from the group consisting of H, (C₁-C₆) alkyl,    heteroaryl and aryl;-   R¹ is selected from the group consisting of halogen, CN, (C₂-C₆)    alkyl, substituted (C₁-C₆) alkyl, aryl, substituted aryl,    heterocyclyl, substituted heterocyclyl and —V—R⁷;-   R² and R³ are selected independently for each occurrence of (CR²R³)    from the group consisting of hydrogen and (C₁-C₆) alkyl;-   R⁴ is selected from a group consisting of alkyl, OH, alkoxy,    heterocyclyl, aryl, substituted alkyl, substituted heterocyclyl, and    substituted aryl;-   R⁷ is chosen from H, (C₁-C₆) alkyl, substituted (C₁-C₆) alkyl, aryl,    substituted aryl, heterocyclyl, and substituted heterocyclyl;-   V is chosen from —C(═O)O—, —C(═O)NR⁸—, —O— and —NR⁸—;-   R⁸ is chosen from H and (C₁-C₆) alkyl, or, when taken together with    the nitrogen to which they are attached, R¹ and R⁸ form a 4-7    membered nitrogen heterocycle;-   R⁹ is chosen from hydrogen, alkyl, and substituted alkyl; and-   y is zero or an integer selected from 1, 2, and 3.

The members of this genus are useful in inhibiting Jak3 activity and assuch are useful in indications where clinical immunosuppression isdesired and in the treatment of hematological cancers.

In another aspect, the invention relates to pharmaceutical compositionscomprising a therapeutically effective amount of at least one compoundof general formula I or II, or a pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier.

In another aspect, the invention relates to a method for treating adisease by altering a response mediated by Jak3 tyrosine kinase. Themethod comprises bringing into contact with Jak3 at least one compoundof general formula I or II.

In yet another aspect the present invention relates to a method ofsuppressing the immune system in a subject in need thereof comprisingadministering to the subject a therapeutically effective amount of atleast one compound of general formula I or II. Suppression of immunesystem activity is desirable for preventing or treating tissue or organrejection following transplant surgery and for preventing and treatingdiseases and disorders arising from aberrant activity of the immunesystem, in particular autoimmune disorders and diseases. Exemplaryautoimmune disorders include graft versus host disease (GVHD),insulin-dependent diabetes (Type I), Hashimoto's thyroiditis and Graves'disease, pernicious anemia, Addison's disease, chronic active hepatitis,Crohn's disease, ulcerative colitis, rheumatoid arthritis, multiplesclerosis, systemic lupus erythematosus, psoriasis, scleroderma andmyasthenia gravis.

The compounds of the present invention are useful in preventing andtreating diseases and disorders related to mast cell-mediated allergicreactions and inflammation.

Other indications in which the Jak3 inhibitors are useful includeleukemias and lymphomas.

DETAILED DESCRIPTION OF THE INVENTION

Throughout this specification the substituents are defined whenintroduced and retain their definitions.

In a first aspect the invention relates to purines and purinones of theformulae:

The members of the genus I may be conveniently divided into twosubgenera based on the values of Q. When Q is nitrogen, a subgenus ofpurinones having an attached imidazo[5,4-c]pyridine arises. When Q iscarbon, a subgenus of purinones having an attached benzimidazole arises.The structures of these subgenera are shown below:

Similarly, the members of the genus II may be conveniently divided intotwo subgenera based on the values of Q. When Q is nitrogen, a subgenusof purines having an attached imidazo[5,4-c]pyridine arises. When Q iscarbon, a subgenus of purines having an attached benzimidazole arises.The structures of these subgenera are shown below:

In certain embodiments, X may be hydrogen, halogen, or anelectron-withdrawing group containing one or fewer carbons. Examplesinclude: H, F, Cl, CN, CF₃, or OCF₃. In some embodiments, y is 1 or 2,and R² and R³ are hydrogen or methyl, and in particular, y may be one,both of R² and R³ may be hydrogen, R⁹ may be hydrogen, and R⁴ may bearyl, heteroaryl, and their substituted counterparts. In otherembodiments, y may be 1 to 3, R² and R³ may be hydrogen in alloccurrences, R⁹ may be hydrogen, and R⁴ may be alkoxy or OH. Theseembodiments have the formulae Ib or IIb:

wherein R⁵ is hydrogen or (C₁-C₆) alkyl.

In yet other embodiments, R⁹ is hydrogen, y is zero, and R⁴ is a residueselected from an optionally substituted monocycle or bicycle. The R⁴residue in this case contains at least one oxygen atom. Moreparticularly, R⁴ may be an oxygen heterocycle, an amide, a substitutedalkyl amide, a halogen-substituted oxygen heterocycle, ahydroxyl-substituted cycloalkyl, a hydroxyl-substituted aryl, or analkoxy-substituted cycloalkyl, such as methoxycyclohexyl, particularlytrans 4-methoxycyclohexyl, or

wherein R⁶ is hydrogen or fluorine. In the chroman, the carbon markedwith an asterisk may be of the R absolute configuration:

In the 4-substituted-1,23,4-tetrahydronaphthalen-1-ol, the carbon atomsmarked with an asterisk may both be of the R absolute configuration:

In other embodiments, A is hydrogen or (C₁-C₆) alkyl. For example, A maybe hydrogen or methyl.

In other embodiments, R¹ is (1) heteroaryl, substituted heteroaryl,heterocyclyl, or substituted heterocyclyl, e.g. pyridinyl, pyrazolyl,pyrimidinyl, isoquinolinyl, azetidinyl, piperidinyl, piperizinyl,pyrrolidinyl, morpholinyl, azepanyl, and diazepanyl; (2) aryl,substituted aryl, (C₂-C₆) alkyl or substituted (C₁-C₆) alkyl; (3)halogen or CN; or (4)-V—R⁷. The foregoing are typically optionallysubstituted with hydroxy, halogen, carboxamide, alkyl, carboxy, sulfone,alkoxy, and cyano.

In the embodiment in which R¹ is VR⁷, V may be —C(═O)O— or —C(═O)NR⁸—and R⁷ may be —CH₂CN, (C₁-C₆) alkyl, or H. Alternatively, VR⁷ may be

Also when R¹ is VR⁷, V may be —O— or —NR⁸—, R⁷ may be (C₁-C₆) alkyl, andR⁸ may be H.

All of the compounds falling within the foregoing parent genera andtheir subgenera are useful as Jak3 inhibitors.

In additional embodiments, R⁹ may be H, Q may be CX and Y may be zero asshown in formulae Ic and IIc:

In some embodiments of formulae Ic and IIc, R¹ is chosen from halogen,(C₂-C₆) alkyl, substituted (C₁-C₆) alkyl, heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, or —V—R⁷ wherein V is —NR⁸—,—C(═O)O—, —C(═O)NR⁸, or O; R⁷ is H, (C₁-C₆) alkyl, substituted (C₁-C₆)alkyl, heterocyclyl, substituted heterocyclyl, or aryl; and R⁸ ishydrogen or (C₁-C₆) alkyl, or when taken together with the nitrogen towhich they are attached, R⁷ and R⁸ form a 4-7 membered nitrogenheterocycle.

Examples of compounds that fall within the scope of the foregoingembodiments include but are not limited to:

In other embodiments R⁴ is chosen from substituted alkyl, heterocyclyl,substituted heterocyclyl, and substituted aryl, e.g. tetrahydrofuranyl,pyranyl, benzopyranyl, hydroxytetralinyl, oxepanyl, hydroxycyclohexyl,and their halogenated congeners; R¹ is chosen from halogen, (C₂-C₆)alkyl, substituted (C₁-C₆) alkyl, phenyl, azetidinyl, piperidinyl,piperizinyl, pyrrolidinyl, morpholinyl, azepanyl, diazepanyl, pyridinyl,pyrimidinyl, and pyrazolyl optionally substituted with hydroxy, halogen,carboxamide, carboxy, sulfone, alkoxy, and cyano; and X is chosen fromhalogen, cyano, and hydrogen.

Examples of compounds that fall within the scope of the foregoingembodiments include but are not limited to:

In a further embodiment, R⁹ may be H, Q may be CX, y may be zero, and R⁴is a hydroxytetralin and hydroxycyclohexyl of formulae Id and IId:

wherein R⁶ is H or halogen. R¹ may be selected from (C₂-C₆) alkyl,substituted (C₁-C₆) alkyl, halogen, azetidinyl, piperidinyl,piperizinyl, pyrrolidinyl, phenyl, morpholinyl, azepanyl, diazepanyl,pyridinyl, pyrimidinyl, and pyrazolyl optionally substituted withhydroxy, halogen, carboxamide, alkyl, carboxy, sulfone, alkoxy, andcyano; and X is halogen, cyano, substituted alkoxy, and hydrogen.

Examples of compounds that are within the scope of the foregoingembodiments include but are not limited to:

In certain embodiments, R⁹ may be alkyl or substituted alkyl, Q may beCX, and y may be zero, and of formula Ie:

In further embodiments, R⁴ may be chosen from tetrahydrofuran,benzopyran, hydroxytetralin, oxepane, hydroxycyclohexane, and theirhalogenated congeners; R¹ may be chosen from halogen, heterocyclyl,substituted heterocyclyl, (C₂-C₆) alkyl, substituted (C₁-C₆) alkyl,aryl, substituted aryl, cyano, carboxy, carboalkoxy, carboxamide, andamidino; and X may be chosen from halogen, cyano, hydrogen, alkoxy, orsubstituted alkoxy.

Compounds that fall within the scope of the foregoing embodimentsinclude but are not limited to:

DEFINITIONS

For convenience and clarity certain terms employed in the specification,examples and claims are described herein.

Alkyl is intended to include linear, branched, or cyclic hydrocarbonstructures and combinations thereof. Lower alkyl refers to alkyl groupsof from 1 to 6 carbon atoms. Examples of lower alkyl groups includemethyl, ethyl, propyl, isopropyl, butyl, s- and t-butyl and the like.Preferred alkyl groups are those of C₂₀ or below; more preferred areC₁-C₈ alkyl. Cycloalkyl is a subset of alkyl and includes cyclichydrocarbon groups of from 3 to 8 carbon atoms. Examples of cycloalkylgroups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,norbornyl, and the like.

C₁ to C₂₀ hydrocarbon includes alkyl, cycloalkyl, alkenyl, alkynyl, aryland combinations thereof. Examples include phenethyl, cyclohexylmethyl,camphoryl and naphthylethyl.

Alkoxy or alkoxyl refers to groups of from 1 to 8 carbon atoms of astraight, branched, cyclic configuration and combinations thereofattached to the parent structure through an oxygen. Examples includemethoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy andthe like. Lower-alkoxy refers to groups containing one to four carbons.The term oxaalkyl is intended as it is understood in the art [see Namingand Indexing of Chemical Substances for Chemical Abstracts, published bythe American Chemical Society, ¶196, but without the restriction of¶127(a)], i.e. it refers to compounds in which the oxygen is bonded viaa single bond to its adjacent atoms (forming ether bonds); it does notrefer to doubly bonded oxygen, as would be found in carbonyl groups.

Acyl refers to groups of from 1 to 8 carbon atoms of a straight,branched, cyclic configuration, saturated, unsaturated and aromatic andcombinations thereof, attached to the parent structure through acarbonyl functionality. One or more carbons in the acyl residue may bereplaced by nitrogen, oxygen or sulfur as long as the point ofattachment to the parent remains at the carbonyl. Examples includeacetyl, benzoyl, propionyl, isobutyryl, t-butoxycarbonyl,benzyloxycarbonyl and the like. Lower-acyl refers to groups containingone to four carbons.

Aryl and heteroaryl mean a 5- or 6-membered aromatic or heteroaromaticring containing 0-3 heteroatoms selected from O, N, or S; a bicyclic 9-or 10-membered aromatic or heteroaromatic ring system containing 0-3heteroatoms selected from O, N, or S; or a tricyclic 13- or 14-memberedaromatic or heteroaromatic ring system containing 0-3 heteroatomsselected from O, N, or S. The aromatic 6- to 14-membered carbocyclicrings include, e.g., benzene and naphthalene, and for the purposes ofthe present invention, fused moieties such as tetrahydronaphthalene(tetralin), indane and fluorine, in which one or more rings arearomatic, but not all need be. The 5- to 10-membered aromaticheterocyclic rings include, e.g., imidazole, pyridine, indole,thiophene, benzopyranone, thiazole, furan, benzimidazole, quinoline,isoquinoline, quinoxaline, pyrimidine, pyrazine, tetrazole and pyrazole.

Arylalkyl refers to a substituent in which an aryl residue is attachedto the parent structure through alkyl. Examples are benzyl, phenethyland the like.

Heteroarylalkyl refers to a substituent in which a heteroaryl residue isattached to the parent structure through alkyl. Examples include, e.g.,pyridinylmethyl, pyrimidinylethyl and the like.

Heterocycle means a cycloalkyl or aryl residue in which from one tothree carbons is replaced by a heteroatom selected from the groupconsisting of N, O and S. The nitrogen and sulfur heteroatoms mayoptionally be oxidized, and the nitrogen heteroatom may optionally bequaternized. Examples of heterocycles include pyrrolidine, pyrazole,pyrrole, indole, quinoline, isoquinoline, tetrahydroisoquinoline,benzofuran, benzodioxan, benzodioxole (commonly referred to asmethylenedioxyphenyl, when occurring as a substituent), tetrazole,morpholine, thiazole, pyridine, pyridazine, pyrimidine, thiophene,furan, oxazole, oxazoline, isoxazole, dioxane, tetrahydrofuran and thelike. It is to be noted that heteroaryl is a subset of heterocycle inwhich the heterocycle is aromatic.

Examples of heterocyclyl residues additionally include piperazinyl,2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxo-pyrrolidinyl, 2-oxoazepinyl,azepinyl, 4-piperidinyl, pyrazolidinyl, imidazolyl, imidazolinyl,imidazolidinyl, pyrazinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl,isothiazolyl, quinuclidinyl, isothiazolidinyl, benzimidazolyl,thiadiazolyl, benzopyranyl, benzothiazolyl, tetrahydrofuryl,tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl,thiamorpholinylsulfoxide, thiamorpholinylsulfone, oxadiazolyl, triazolyland tetrahydroquinolinyl. A nitrogenous heterocycle is a heterocyclecontaining at least one nitrogen in the ring; it may contain additionalnitrogens, as well as other heteroatoms.

Substituted alkyl, aryl, cycloalkyl, heterocyclyl etc. refer to alkyl,aryl, cycloalkyl, or heterocyclyl wherein up to three H atoms in eachresidue are replaced with halogen, haloalkyl, hydroxy, loweralkoxy,hydroxyloweralkyl, carboxy, carboalkoxy (also referred to asalkoxycarbonyl), carboxamido (also referred to as alkylaminocarbonyl),cyano, carbonyl, nitro, amino, alkylamino, dialkylamino, mercapto,alkylthio, sulfoxide, sulfone, acylamino, amidino, phenyl, benzyl,heteroaryl, phenoxy, benzenesulfonyl, benzyloxy, or heteroaryloxy. Whenthe parent is a heterocycle that allows such substitution, the term alsoincludes oxides, for example pyridine-N-oxide, thiopyran sulfoxide andthiopyran-S,S-dioxide. As mentioned above, two hydrogens on a singlecarbon may be replaced by a carbonyl to form an oxo derivative.Noteworthy oxo-substituted aryl residues include tetralone(3,4-dihydronaphthalen-1(2H)-one) and indanone (2,3-dihydroinden-1-one).

The terms “halogen” and “halo” refer to fluorine, chlorine, bromine oriodine.

The term “electron-withdrawing group” refers to substituents which havea Hammett σ_(meta) greater than 0.2. Examples of such substituentsinclude cyanide, trifluoromethoxy, trifluoromethyl, chlorine, andfluorine.

Some of the compounds described herein may contain one or moreasymmetric centers and may thus give rise to enantiomers, diastereomers,and other stereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-. The present invention is meant toinclude all such possible isomers, as well as, their racemic andoptically pure forms. Optically active (R)- and (S)-isomers may beprepared using chiral synthons or chiral reagents, or resolved usingconventional techniques. When the compounds described herein containolefinic double bonds or other centers of geometric asymmetry, andunless specified otherwise, it is intended that the compounds includeboth E and Z geometric isomers.

Likewise, all tautomeric forms are also intended to be included. Whenthe enol-form of a tautomeric compound is present, the keto-form of thecompound is also within the scope of the present invention. An exampleof a keto-enol tautomerization of a 6-substituted 2-(benzimidazolyl)purinone and subsequently a tautomer of the foregoing that is inaccordance with the present invention are depicted below.

The graphic representations of racemic, ambiscalemic and scalemic orenantiomerically pure compounds used herein are taken from Maehr J.Chem. Ed. 62, 114-120 (1985): solid and broken wedges are used to denotethe absolute configuration of a chiral element; wavy lines indicatedisavowal of any stereochemical implication which the bond it representscould generate; solid and broken bold lines are geometric descriptorsindicating the relative configuration shown; and wedge outlines anddotted or broken lines denote enantiomerically pure compounds ofindeterminate absolute configuration. For example, the graphicrepresentation

indicates either, or both, of the two trans enantiomers

It will be recognized that the compounds of this invention can exist inradiolabeled form, i.e., the compounds may contain one or more atomscontaining an atomic mass or mass number different from the atomic massor mass number usually found in nature. Radioisotopes of hydrogen,carbon, phosphorous, fluorine, chlorine and iodine include ³H, ¹⁴C, ³⁵S,¹⁸F, ³⁶Cl and ¹²⁵I, respectively. Compounds that contain thoseradioisotopes and/or other radioisotopes of other atoms are within thescope of this invention. Tritiated, i.e. ³H, and carbon-14, i.e., ¹⁴C,radioisotopes are particularly preferred for their ease in preparationand detectability. Radiolabeled compounds of this invention cangenerally be prepared by methods well known to those skilled in the art.Conveniently, such radiolabeled compounds can be prepared by carryingout the procedures disclosed in the Examples by substituting a readilyavailable radiolabeled reagent for a non-radiolabeled reagent. Becauseof the high affinity for the JAK3 enzyme active site, radiolabeledcompounds of the invention are useful for JAK3 assays.

An oxygenous heterocycle is a heterocycle containing at least one oxygenin the ring; it may contain additional oxygens, as well as otherheteroatoms. Exemplary oxygenous heterocycles include tetrahydropyran,chroman and their variously substituted derivatives, such as:

Chemical Synthesis

Terminology related to “protecting”, “deprotecting” and “protected”functionalities occurs throughout this application. Such terminology iswell understood by persons of skill in the art and is used in thecontext of processes that involve sequential treatment with a series ofreagents. In that context, a protecting group refers to a group which isused to mask a functionality during a process step in which it wouldotherwise react, but in which reaction is undesirable. The protectinggroup prevents reaction at that step, but may be subsequently removed toexpose the original functionality. The removal or “deprotection” occursafter the completion of the reaction or reactions in which thefunctionality would interfere. Thus, when a sequence of reagents isspecified, as it is in the processes of the invention, the person ofordinary skill can readily envision those groups that would be suitableas “protecting groups”. Suitable groups for that purpose are discussedin standard textbooks in the field of chemistry, such as ProtectiveGroups in Organic Synthesis by T. W. Greene [John Wiley & Sons, NewYork, 1991], which is incorporated herein by reference.

A comprehensive list of abbreviations utilized by organic chemistsappears in the first issue of each volume of the Journal of OrganicChemistry. The list, which is typically presented in a table entitled“Standard List of Abbreviations”, is incorporated herein by reference.

In general, the compounds of the present invention may be prepared bythe methods illustrated in the general reaction schemes as, for example,described below, or by modifications thereof, using readily availablestarting materials, reagents and conventional synthesis procedures. Inthese reactions, it is also possible to make use of variants that are inthemselves known, but are not mentioned here. The starting materials,for example in the case of suitably substituted benzimidazole ringcompounds, are either commercially available, synthesized as describedin the examples or may be obtained by the methods well known to personsof skill in the art.

The present invention further provides pharmaceutical compositionscomprising as active agents, the compounds described herein.

As used herein a “pharmaceutical composition” refers to a preparation ofone or more of the compounds described herein, or physiologicallyacceptable salts or solvents thereof, with other chemical componentssuch as physiologically suitable carriers and excipients. Pharmaceuticalcompositions for use in accordance with the present invention thus maybe formulated in conventional manner using one or more physiologicallyacceptable carriers comprising excipients and auxiliaries, whichfacilitate processing of the active compounds into preparations which,can be used pharmaceutically. Proper formulation is dependent upon theroute of administration chosen.

Compounds that inhibit Jak-3 can be formulated as pharmaceuticalcompositions and administered to a mammalian subject, such as a humanpatient in a variety of forms adapted to the chosen route ofadministration, i.e., orally or parenterally, by intravenous,intramuscular, topical, transdermal or subcutaneous routes.

For oral administration, the compounds can be formulated readily bycombining the active compounds with pharmaceutically acceptable carrierswell known in the art. Such carriers enable the compounds of theinvention to be formulated as tablets, pills, dragees, capsules,liquids, gels, syrups, slurries, suspensions, and the like, for oralingestion by a patient. Pharmacological preparations for oral use can bemade using a solid excipient, optionally grinding the resulting mixture,and processing the mixture of granules, after adding suitableauxiliaries if desired, to obtain tablets or dragee cores. Suitableexcipients are, in particular, fillers such as sugars, includinglactose, sucrose, mannitol, or sorbitol; cellulose preparations such as,for example, maize starch, wheat starch, rice starch, potato starch,gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carbomethylcellulose; and/orphysiologically acceptable polymers such as polyvinylpyrrolidone (PVP).If desired, disintegrating agents may be added, such as cross-linkedpolyvinyl pyrrolidone, agar or alginic acid or a salt thereof such assodium alginate.

In addition, enteric coating may be useful as it is may be desirable toprevent exposure of the compounds of the invention to the gastricenvironment. Pharmaceutical compositions, which can be used orally,include push-fit capsules made of gelatin as well as soft, sealedcapsules made of gelatin and a plasticizer, such as glycerol orsorbitol. The push-fit capsules may contain the active ingredients inadmixture with filler such as lactose, binders such as starches,lubricants such as talc or magnesium stearate and, optionally,stabilizers.

In soft capsules, the active compounds may be dissolved or suspended insuitable liquids, such as fatty oils, liquid paraffin, or liquidpolyethylene glycols. In addition, stabilizers may be added. Allformulations for oral administration should be in dosages suitable forthe chosen route of administration.

For injection, the compounds of the invention may be formulated inaqueous solutions, preferably in physiologically compatible buffers suchas Hank's or Ringer's solution or physiological saline buffer. Fortransmucosal and transdermal administration, penetrants appropriate tothe barrier to be permeated may be used in the composition. Suchpenetrants, including for example DMSO or polyethylene glycol, are knownin the art.

For administration by inhalation, the compounds for use according to thepresent invention are conveniently delivered in the form of an aerosolspray presentation from a pressurized pack or a nebulizer with the useof a suitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichloro-tetrafluoroethane or carbon dioxide. Inthe case of a pressurized aerosol, the dosage unit may be determined byproviding a valve to deliver a metered amount. Capsules and cartridgesof, e.g., gelatin for use in an inhaler or insufflator may be formulatedcontaining a powder mix of the compound and a suitable powder base suchas lactose or starch.

Pharmaceutical compositions for parenteral administration includeaqueous solutions of the active ingredients in water-soluble form.Additionally, suspensions of the active compounds may be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidsesters such as ethyl oleate, triglycerides or liposomes. Aqueousinjection suspensions may contain substances, which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol or dextran. Optionally, the suspension may also containsuitable stabilizers or agents, which increase the solubility of thecompounds, to allow for the preparation of highly concentratedsolutions.

The compounds of the present invention may also be formulated in rectalcompositions such as suppositories or retention enemas, using, e.g.,conventional suppository bases such as cocoa butter or other glycerides.

Depending on the severity and responsiveness of the condition to betreated, dosing can also be a single administration of a slow releasecomposition, with course of treatment lasting from several days toseveral weeks or until cure is effected or diminution of the diseasestate is achieved. The amount of a composition to be administered will,of course, be dependent on many factors including the subject beingtreated, the severity of the affliction, the manner of administration,the judgment of the prescribing physician. The compounds of theinvention may be administered orally or via injection at a dose from0.001 to 2500 mg/kg per day. The dose range for adult humans isgenerally from 0.005 mg to 10 g/day. Tablets or other forms ofpresentation provided in discrete units may conveniently contain anamount of compound of the invention which is effective at such dosage oras a multiple of the same, for instance, units containing 5 mg to 500mg, usually around 10 mg to 200 mg. The precise amount of compoundadministered to a patient will be the responsibility of the attendantphysician. However, the dose employed will depend on a number offactors, including the age and sex of the patient, the precise disorderbeing treated, and its severity. Also, the route of administration mayvary depending on the condition and its severity.

As used herein, and as would be understood by the person of skill in theart, the recitation of “a compound” is intended to include salts,solvates and inclusion complexes of that compound. The term “solvate”refers to a compound of Formula I or II in the solid state, whereinmolecules of a suitable solvent are incorporated in the crystal lattice.A suitable solvent for therapeutic administration is physiologicallytolerable at the dosage administered. Examples of suitable solvents fortherapeutic administration are ethanol and water. When water is thesolvent, the solvate is referred to as a hydrate. In general, solvatesare formed by dissolving the compound in the appropriate solvent andisolating the solvate by cooling or using an antisolvent. The solvate istypically dried or azeotroped under ambient conditions. Inclusioncomplexes are described in Remington: The Science and Practice ofPharmacy 19th Ed. (1995) volume 1, page 176-177, which is incorporatedherein by reference. The most commonly employed inclusion complexes arethose with cyclodextrins, and all cyclodextrin complexes, natural andsynthetic, are specifically encompassed within the claims.

The term “pharmaceutically acceptable salt” refers to salts preparedfrom pharmaceutically acceptable non-toxic acids or bases includinginorganic acids and bases and organic acids and bases. When thecompounds of the present invention are basic, salts may be prepared frompharmaceutically acceptable non-toxic acids including inorganic andorganic acids. Suitable pharmaceutically acceptable acid addition saltsfor the compounds of the present invention include acetic,benzenesulfonic (besylate), benzoic, camphorsulfonic, citric,ethenesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric,isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic,nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaricacid, p-toluenesulfonic, and the like. When the compounds contain anacidic side chain, suitable pharmaceutically acceptable base additionsalts for the compounds of the present invention include metallic saltsmade from aluminum, calcium, lithium, magnesium, potassium, sodium andzinc or organic salts made from lysine, N,N′-dibenzylethylenediamine,chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine(N-methylglucamine) and procaine.

The term “preventing” as used herein refers to administering amedicament beforehand to forestall or obtund an attack. The person ofordinary skill in the medical art (to which the present method claimsare directed) recognizes that the term “prevent” is not an absoluteterm. In the medical art it is understood to refer to the prophylacticadministration of a drug to substantially diminish the likelihood orseriousness of a condition, and this is the sense intended herein.

It should be understood that in addition to the ingredients particularlymentioned above, the formulations of this invention may include otheragents conventional in the art having regard to the type of formulationin question, for example those suitable for oral administration mayinclude flavoring agents.

The compositions may be presented in a packaging device or dispenser,which may contain one or more unit dosage forms containing the activeingredient. Examples of a packaging device include metal or plasticfoil, such as a blister pack and a nebulizer for inhalation. Thepackaging device or dispenser may be accompanied by instructions foradministration. Compositions comprising a compound of the presentinvention formulated in a compatible pharmaceutical carrier may also beplaced in an appropriate container and labeled for treatment of anindicated condition.

Indications

The compounds of the present invention are useful in inhibiting theactivity if Jak3 or in inhibiting Jak3 mediated activity and are usefulas immunosuppressive agents for tissue and organ transplants, includingbone marrow transplant and in the treatment of autoimmune andinflammatory diseases and of complications arising therefrom.Hyperacute, acute and chronic organ transplant rejection may be treated.Hyperacute rejection occurs within minutes of transplantation. Acuterejection generally occurs within six to twelve months of thetransplant. Hyperacute and acute rejections are typically reversiblewhere treated with immunosuppressant agents. Chronic rejection,characterized by gradual loss of organ function, is an ongoing concernfor transplant recipients because it can occur anytime aftertransplantation.

There are about 75 different autoimmune disorders known that may beclassified into two types, organ-specific (directed mainly at one organ)and non-organ-specific (affecting multiple organs).

Examples of organ-specific autoimmune disorders are insulin-dependentdiabetes (Type I) which affects the pancreas, Hashimoto's thyroiditisand Graves' disease which affect the thyroid gland, pernicious anemiawhich affects the stomach, Cushing's disease and Addison's disease whichaffect the adrenal glands, chronic active hepatitis which affects theliver; polycystic ovary syndrome (PCOS), celiac disease, psoriasis,inflammatory bowel disease (IBD) and ankylosing spondylitis.

Examples of non-organ-specific autoimmune disorders are rheumatoidarthritis, multiple sclerosis, systemic lupus and myasthenia gravis.

Type I diabetes ensues from the selective aggression of autoreactiveT-cells against insulin secreting β cells of the islets of Langerhans.Targeting Jak3 in this disease is based on the observation that multiplecytokines that signal through the Jak pathway are known to participatein the T-cell mediated autoimmune destruction of β cells. Indeed, a Jak3inhibitor, JANEX-1 was shown to prevent spontaneous autoimmune diabetesdevelopment in the NOD mouse model of type I diabetes.

Graft-versus-host disease (GVHD) is a donor T-cell initiatedpathological condition that frequently follows allogeneic bone marrowtransplantation (BMT). Substantial experimental and clinical researchhave demonstrated that donor T-cells are the principal mediators andeffectors of GVHD. Jak3 plays a key role in the induction of GVHD andtreatment with a Jak3 inhibitor, JANEX-1, was shown to attenuate theseverity of GVHD (reviewed in Cetkovic-Cvrlje and Ucken, 2004).

Mast cells express Jak3 and Jak3 is a key regulator of the IgE mediatedmast cell responses including the release of inflammatory mediators.Jak3 was shown to be a valid target in the treatment of mast cellmediated allergic reaction.

Allergic disorders associated with mast cell activation include Type Iimmediate hypersensitivity reactions such as allergic rhinitis (hayfever), allergic urticaria (hives), angioedema, allergic asthma andanaphylaxis, i.e., “anaphylatic shock.” These disorders are treated orprevented by inhibition of Jak3 activity, for example, by administrationof a Jak3 inhibitor according to the present invention. According to thepresent invention, the Jak3 inhibitors may be administeredprophylactically, i.e., prior to onset of acute allergic reaction, orthey may be administered after onset of the reaction, or at both times.

Inflammation of tissues and organs occurs in a wide range of disordersand diseases and in certain variations, results from activation of thecytokine family of receptors. Exemplary inflammatory disordersassociated with activation of Jak3 include, in a non-limiting manner,skin inflammation due radiation exposure, asthma, allergic inflammationand chronic inflammation.

The Jak3 inhibitors of the present invention are also useful in treatingcertain malignancies, including skin cancer and hematological malignancysuch as lymphomas and leukemias.

The following examples will further describe the invention, and are usedfor the purposes of illustration only, and should not be considered aslimiting the invention being disclosed.

EXAMPLES

The following abbreviations and terms have the indicated meaningthroughout:

-   -   Ac=acetyl    -   Bu=butyl    -   DCM=dichloromethane=methylene chloride=CH₂Cl₂    -   DEAD=diethyl azodicarboxylate    -   DIC=diisopropylcarbodiimide    -   DIEA=N,N-diisopropylethyl amine    -   DMF=N,N-dimethylformamide    -   DMSO=dimethyl sulfoxide    -   EA (EtOAc)=Ethyl Acetate    -   GC=gas chromatography    -   h=hours    -   HOAc=acetic acid    -   HOBt=hydroxybenzotriazole    -   Me=methyl    -   Pd(dppf)₂Cl₂=dichloro[1,1′-bis(diphenylphosphinoferrocene]palladium    -   Ph=phenyl    -   PhOH=phenol    -   RT=room temperature    -   sat'd=saturated    -   s-=secondary    -   t-=tertiary    -   TBDMS=t-butyldimethylsilyl    -   TFA=trifluoroacetic acid    -   THF=tetrahydrofuran    -   TMOF=trimethyl orthoformate    -   TMS=trimethylsilyl    -   tosyl=p-toluenesulfonyl    -   Trt=triphenylmethyl

Examples below describe syntheses of compounds, precursors andintermediates of the invention.

Experimental Part

6-chloro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine.2,5-diamino-4,6-dichloropyrimidine (6 g, 33.5 mmol),4-aminotetrahydropyran (3.39 g, 33.5 mmol, 1 equiv.), sodium bicarbonate(9.85 g, 117.2 mmol, 3.5 equiv.) and 1-butanol (120 mL) were heatedtogether at 150° C. in a sealed tube. After 3 days, when the reactionappeared to be complete (by HPLC), the reaction mixture was cooled toroom temperature and the solvent was removed in vacuo. The residue waspurified by flash chromatography (silica gel, gradual elution with 95/5methylene chloride/methanol to 90/10 methylene chloride/methanol) togive 7.1 g (87% yield) of the desired product as a pink solid. In analternative procedure, water (300 mL) was added to the residue (beforechromatographic purification) and the mixture stirred for 30 min at roomtemperature. Filtration under vacuum, thorough washing of theprecipitate with water, followed by thorough drying of the pink solid invacuum oven at 60° C., provided high purity desired product in 78%yield. ¹H NMR (300 MHz, CD₃OD) δ, ppm: 4.21-4.12 (m, 1H), 4.01-3.96 (m,2H), 3.53 (td, J=11.7, 2.2 Hz, 2H), 2.01-1.96 (m, 2H), 1.60 (tdd,J=12.1, 11.7, 4.6 Hz, 2H); MS (EI) m/z 244.3 (MH)⁺.

2-amino-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one. To asolution of6-chloro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine (1 g,4.1 mmol) in 900 mL of anhydrous THF at −78° C. under Ar was addeddropwise, over 40 min, a solution of phosgene in THF (3.8 mL of a 20%solution of phosgene in toluene, 7 mmol, 1.7 equiv., diluted with 26 mLof anhydrous THF). The reaction mixture was left to gradually warm up toroom temperature over 16 h. It was purged with air for 30 min, then thesolvent was removed in vacuo to give 1.2 g (97% yield) of the desiredproduct (HCl salt) as a white solid. ¹H NMR (300 MHz, d₆-DMSO) δ, ppm:11.40 (s, 1H), 7.00 (br s, 2H), 4.50-4.37 (m, 1H), 4.12-4.04 (m, 2H),3.50 (app t, 2H), 2.68-2.55 (m, 2H), 1.78-1.71 (m, 2H); MS (EI) m/z270.3 (MH)⁺.

Alternative route to2-amino-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one. To asolution of6-chloro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine (5.1 g,20.9 mmol) in 170 mL of anhydrous THF was added 1,1-carbonyldiimidazole(20.4 g, 126 mmol) as a solid, in portions. The reaction mixture wasstirred at room temperature for 20-30 min (completion of the reactionchecked by HPLC and MS). The solvent was removed in vacuo. Water (250mL) was added to the residue and the mixture stirred at room temperaturefor 10 min. The solid formed was filtered under vacuum, and thoroughlydried to give 4.67 g (83% yield) of the desired product as a pink solid.¹H NMR (300 MHz, d₆-DMSO) δ, ppm: 11.40 (s, 1H), 7.00 (br s, 2H),4.50-4.37 (m, 1H), 4.12-4.04 (m, 2H), 3.50 (app t, 2H), 2.68-2.55 (m,2H), 1.78-1.71 (m, 2H); MS (EI) m/z 270.3 (MH)⁺.

General procedure for the Buchwald-Hartwig Palladium-catalyzed C—N crosscoupling. All glassware was dried in vacuum oven at 60° C. for one dayprior to reaction. Cesium carbonate and2-amino-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one weredried at 60° C. under high vacuum for one day prior to experiment. Anoven-dried vial was charged with2-amino-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one (1equiv.), freshly grounded cesium carbonate (1.4 equiv.), Pd(OAc)₂ (0.1equiv.), racemic BINAP (0.15 equiv.) and an aryl bromide (1.3 equiv.),followed by anhydrous toluene (0.25 M solution). The vial was purgedwith Ar for 3 min, then closed and heated at 80-100° C. for 17-22 h. Thereaction mixture was cooled to room temperature, and the solvent removedin vacuo. Column chromatography of the residue afforded the desiredproduct.

6-chloro-2-(5-fluoro-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.To an oven-dried vial was added2-amino-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one (50 mgof HCl salt, 0.186 mmol, 1 equiv.) in anhydrous toluene (3 mL), thenfreshly grounded cesium carbonate (85 mg, 0.26 mmol, 1.4 equiv.) withstirring at room temperature under Ar. After 20 min, Pd(OAc)₂ (12.5 mg,0.019 mmol, 0.1 equiv.), racemic BINAP (17.3 mg, 0.028 mmol, 0.15equiv.) and 1-bromo-5-fluoro-2-nitrobenzene (Oakwood) (53 mg, 0.24 mmol,1.3 equiv.) were added as solids, followed by DIEA (50 μL, 1.5 equiv.).The vial was purged with Ar for 3 min, then closed and heated at 80° C.for 18 h. The reaction mixture was cooled to room temperature, dilutedwith 5% methanol in methylene chloride, filtered through a Nylon 0.45 μmfilter, and the filter repeatedly washed with 5% methanol in methylenechloride. The filtrate was concentrated in vacuo and the resultingresidue was purified using preparative TLC (silica gel, 4.25% methanolin methylene chloride) to give the desired product as a yellow solid(45.3 mg, 60% yield). ¹H NMR (300 MHz, CDCl₃) δ, ppm: 10.90 (s, 1H),8.89 (dd, 1H), 8.45 (dd, 1H), 8.10 (br s, 1H), 6.92-6.83 (m, 1H),4.75-4.60 (m, 1H), 4.28 (dd, 2H), 3.65 (app t, 2H), 2.82 (tdd, 2H), 1.90(br d, 2H); MS (EI) m/z 409.1 (MH)⁺.

6-chloro-2-(5-chloro-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.To an oven-dried 2-neck flask under Ar was added racemic BINAP (2.27 g,3.64 mmol, 0.35 equiv.) followed by anhydrous toluene (20 mL) andPd(OAc)₂ (1.05 g, 1.56 mmol, 0.15 equiv.) and the mixture was stirredunder Ar, at room temperature for 15 min. A deep yellow paste is formed.Then, 2-bromo-4-chloro-1 nitrobenzene (3.19 g, 13.52 mmol, 1.3 equiv.),2-amino-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one (2.80g, 10.4 mmol, 1.0 equiv.) and cesium carbonate (5.08 g, 15.6 mmol, 1.5equiv.) were added as solids, under Ar, followed by anhydrous toluene(22 mL), and the reaction mixture was stirred at room temperature for 10min, then stirred at 100° C. for 17 h. The reaction mixture was cooledto room temperature, diluted with toluene (450 mL) and passed throughCelite. The filtrate was concentrated in vacuo to give a dark brownresidue. Flash chromatographic purification (silica gel, gradual elutionwith 1 to 5% methanol in methylene chloride), followed byrecrystallization from hot ethyl acetate gave the desired product as abright yellow solid (1.63 g, 37% yield) in high purity. ¹H NMR (300 MHz,CDCl₃+CD₃OD) δ, ppm: 9.05 (s, 1H), 8.24 (d, 1H), 7.06 (d, 1H), 4.56-4.52(m, 1H), 4.16 (dd, 2H), 3.58 (app t, 2H), 2.72 (tdd, 2H), 1.81 (br d,2H); MS (EI) m/z 425.2 (MH)⁺.

3-(6-chloro-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-ylamino)-4-nitrobenzonitrile.To an oven-dried 2-neck flask under Ar was added racemic BINAP (2.59 g,4.17 mmol, 0.35 equiv.) followed by anhydrous toluene (20 mL) andPd(OAc)₂ (1.20 g, 1.79 mmol, 0.15 equiv.) and the mixture was stirredunder Ar, at room temperature for 15 min. A deep yellow paste is formed.Then, 3-bromo-4-nitro-benzonitrile (3.51 g, 15.47 mmol, 1.3 equiv.),2-amino-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one (3.2 g,11.9 mmol, 1.0 equiv.) and cesium carbonate (5.81 g, 17.8 mmol, 1.5equiv.) were added as solids, under Ar, followed by anhydrous toluene(27 mL), and the reaction mixture was stirred at room temperature for 10min, then stirred at 100° C. for 22 h (LC-MS shows reaction to becomplete). The reaction mixture was cooled to room temperature, dilutedwith toluene (450 mL) and passed through Celite. The filtrate wasconcentrated in vacuo to give a dark brown residue. The desired productwas recrystallized as a bright orange solid from the residue usingmixtures of 100 mL acetonitrile/100 mL water. Repeatedrecrystallizations resulted in isolation of 1.49 g of desired product(30% yield) in high purity. ¹H NMR (300 MHz, CDCl₃) δ, ppm: 10.40 (s,1H), 9.33 (s, 1H), 8.35 (d, 1H), 7.85 (br s, 1H), 7.31 (d, 1H),4.65-4.50 (m, 1H), 4.19 (dd, 2H), 3.55 (app t, 2H), 2.70 (tdd, 2H), 1.90(br d, 2H); MS (EI) m/z 416.2 (MH)⁺.

General procedure for the reduction of the nitrophenyl group in6-chloro-2-(5-substituted-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-onewith iron powder. To6-chloro-2-(5-substituted-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(1 equiv.) was added glacial acetic acid, water and ethanol (ratio1/2.5/5 vol), followed by iron powder (10 equiv.) and the resultingmixture heated at 90° C. for 15 min (HPLC monitoring). The reactionmixture was left to cool down to room temperature, concentrated ammoniumhydroxide solution was added to bring the pH to basic, and the mixturewas stirred for 10 min. The aqueous layer diluted with water, wasextracted with ethyl acetate, the combined organic layers were washedwith brine, dried (anhydrous Na₂SO₄), and the solvent removed in vacuoto give the desired product. This material was used in the next stepwithout further purification.

2-(2-amino-5-chlorophenylamino)-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.Pale yellow solid. Synthesized from6-chloro-2-(5-chloro-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one,using glacial acetic acid, water and ethanol (57 mL/142 mL/285 mL). MS(EI) m/z 395.2 (MH)⁺.

General procedure for the reduction of the nitrophenyl group in6-chloro-2-(5-substituted-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-oneby catalytic hydrogenation. To6-chloro-2-(5-substituted-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(1 equiv.) was added ethyl acetate and Pt catalyst, 0.5% sulfur (AlfaAesar) (5% on activated carbon, 0.05 equiv.) with stirring at roomtemperature under hydrogen (1 atm) for 18 h (HPLC monitoring).Filtration of the reaction mixture over a small plug of Celite, thoroughwashing with ethyl acetate and methanol, evaporation of solvent anddrying provided the desired product, which was used in the next stepwithout further purification.

2-(2-amino-5-fluorophenylamino)-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.160 mg on 0.49 mmol scale, 86% yield. Synthesized from6-chloro-2-(5-fluoro-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.MS (EI) m/z 379.3 (MH)⁺.

4-amino-3-(6-chloro-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-ylamino)benzonitrile.Pale yellow solid. 86% yield on 0.49 mmol scale. In a typical procedure,to3-(6-chloro-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-ylamino)-4-nitrobenzonitrile(0.49 mmol) was added ethyl acetate (20 mL) and Pt catalyst, 0.5% sulfur(Alfa Aesar) (95.5 mg 5% on activated carbon, 4.8 mg, 0.025 mmol, 0.05equiv.) with stirring at room temperature under hydrogen (1 atm) for 18h (HPLC monitoring). Filtration of the reaction mixture over a smallplug of celite, thorough washing with ethyl acetate and methanol,evaporation of solvent and drying provided the desired product, whichwas used in the next step without further purification. Yieldquantitative on 2.95 mmol scale. The catalyst was washed with 12-15portions of 100 mL of a 1/1 (v) solution of methylenechloride/isopropanol to recover desired product. MS (EI) m/z 386.1(MH)⁺.

General procedure for closing the benzimidazole ring. To a flaskcontaining crude2-(2-amino-5-substituted-phenylamino)-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(1 equiv.) was added anhydrous methanol, followed by anhydroustrimethylorthoformate (excess) and methane sulfonic acid (catalyticamount) and the reaction mixture was stirred under Ar at roomtemperature for 2 h (HPLC monitoring). The solvent was removed in vacuoand the residue purified by column chromatography to afford the desiredproduct.

6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.Pale yellow solid. Synthesized from2-(2-amino-5-fluorophenylamino)-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(160 mg, 0.42 mmol) in anhydrous methanol (10 mL), using anhydroustrimethylorthoformate (1 mL) and methane sulfonic acid (5 drops) withstirring under Ar at room temperature for 2 h (HPLC monitoring). Thesolvent was removed in vacuo and the residue purified by columnchromatography (silica gel, gradual elution with 2% methanol inmethylene chloride to 10% methanol in methylene chloride) to afford thedesired product (120 mg, 73% yield). ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ,ppm: 9.03 (s, 1H), 8.40-8.20 (m, 1H), 7.82-7.72 (m, 1H), 7.17-7.11 (m,1H), 4.72-4.58 (m, 1H), 4.28-4.13 (m, 2H), 3.70-3.62 (m, 2H), 2.90-2.72(m, 2H), 1.90-1.78 (m, 2H); MS (EI) m/z 389.2 (MH)⁺.

6-chloro-2-(6-chloro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.Pale yellow solid. 46% yield on 2 steps (reduction and benzimidazoleclosing), on 2.47 mmol scale. Synthesized from2-(2-amino-5-chlorophenylamino)-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.08 (s, 1H), 8.61 (s, 1H), 7.74(d, 1H), 7.40 (d, 1H), 4.65-4.58 (m, 1H), 4.23-4.19 (m, 2H), 3.62 (appt,2H), 2.82-2.76 (m, 2H), 1.93 (br d, 2H); MS (EI) m/z 405.2 (MH)⁺.

3-(6-chloro-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.Pale yellow solid. 73% yield (0.42 mmol scale). Synthesized from4-amino-3-(6-chloro-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-ylamino)benzonitrile.¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.27 (s, 1H), 8.99 (s, 1H), 7.93(d, 1H), 7.71 (d, 1H), 4.72-4.58 (m, 1H), 4.23-4.19 (m, 2H), 3.64-3.62(m, 2H), 2.82-2.78 (m, 2H), 1.90-1.85 (m, 2H); MS (EI) m/z 396.2 (MH)⁺.

Method 1

General procedure for cross-coupling reactions of 6-chloropurinones with(hetero)arylboronic acids. Ethanol (2 mL) was added to an argon-purgedvial containing6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(25 mg, 0.064 mmol), (hetero)aryl boronic acid (0.128 mmol, 2 equiv.),Pd(PPh₃)₄ (7 mg, 0.006 mmol, 0.1 equiv.) and a 2M aqueous solution ofNa₂CO₃ (200 μL), and the mixture was heated for 30 min in the microwaveoven at 150° C. After cooling to ambient temperature, the reactionmixture was diluted with ethanol, filtered through a Nylon 0.45 μmfilter and the filtrate concentrated in vacuo. Preparative HPLCpurification of the residue afforded, after evaporation and drying, thedesired compound.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.White solid. TFA salt. 56% yield. Prepared from pyridine-4-yl boronicacid. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.22 (br s, 1H), 8.97 (br m,2H), 8.37 (d, 1H), 8.22 (br m, 2H), 7.91-7.82 (m, 1H), 7.28-7.19 (m,1H), 4.83-4.71 (m, 1H), 4.35-4.23 (m, 2H), 3.69 (app t, 2H), 3.01-2.82(m, 2H), 1.92 (br d, 2H); MS (EI) m/z 432.2 (MH)⁺.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-6-(1H-pyrazol-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.TFA salt. 10% yield. Prepared from 1H-pyrazole-4-boronic acid. ¹H NMR(300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.22 (br s, 1H), 8.47-8.30 (m, 3H),7.83-7.72 (m, 1H), 7.23-7.11 (m, 1H), 4.73-4.61 (m, 1H), 4.25-4.13 (m,2H), 3.61 (app t, 2H), 2.92-2.73 (m, 2H), 1.83 (br d, 2H); MS (EI) m/z421.4 (MH)⁺.

Method 2

General procedure for displacement reactions of 6-chloropurinones withaliphatic benzylic amine. To 0.049 mmol of6-chloro-2-(6-substituted-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-onedissolved in 1 mL of DMSO was added 1 mL of 1-butanol, followed by analiphatic/benzylic amine (0.2 mmol, 4 equiv.), and the reaction mixturewas stirred at 110° C. for 18 h (HPLC monitoring). Upon completion ofthe reaction, the mixture was left to cool to room temperature, wasdiluted with acetonitrile and filtered through a Nylon 0.45 μm filter.Preparative HPLC purification afforded, after solvent evaporation anddrying, the desired product.

3-(6-(azetidin-1-yl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.White solid. ¹H NMR (300 MHz, d₆-DMSO) δ, ppm: 11.13 (br s, 1H), 9.24(s, 1H), 8.86 (d, J=0.9 Hz, 1H), 7.96 (d, J=8.4 Hz, 1H), 7.75 (dd, 1H),4.60-4.40 (m, 1H), 4.34 (t, 4H), 4.05-4.00 (dd, 2H), 3.51-3.38 (dd, 2H),2.61-2.42 (m, 4H), 1.72 (br d, 2H); MS (EI) m/z 417.1 (MH)⁺.

3-(6-(3,3-difluoroazetidin-1-yl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.White solid. TFA salt. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.17 (s,1H), 8.87 (br s, 1H), 7.88 (d, 1H), 7.64 (dd, 1H), 4.71 (t, 4H),4.62-4.52 (m, 1H), 4.17 (dd, 2H), 3.58 (app t, 2H), 2.84-2.68 (m, 2H),1.79 (br d, 2H); MS (EI) m/z 453.1 (MH)⁺.

3-(6-(cyclopropylmethylamino)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.Pale yellow solid. TFA salt. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.23(s, 1H), 9.01 (s, 1H), 7.88 (d, 1H), 7.63 (dd, 1H), 4.62-4.44 (m, 1H),4.18 (dd, 2H), 3.60 (app t, 2H), 3.52 (d, 2H), 2.87-2.73 (m, 2H), 1.83(br d, 2H), 1.26-1.20 (m, 1H), 0.67-0.61 (dd, 2H), 0.37 (d, 2H); MS (EI)m/z 431.2 (MH)⁺.

3-(6-morpholino-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.White solid. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.19 (s, 1H), 8.91(s, 1H), 7.89 (d, 1H), 7.65 (d, 1H), 4.72-4.58 (m, 1H), 4.19 (dd, 2H),3.91-3.87 (m, 4H), 3.79-3.75 (m, 4H), 3.60 (app t, 2H), 2.99-2.78 (m,2H), 1.83 (br d, 2H); MS (EI) m/z 447.1 (MH)⁺.

2-(6-chloro-1H-benzo[d]imidazol-1-yl)-6-(3-hydroxyazetidin-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.Pale yellow solid (TFA salt). ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.17(s, 1H), 8.68 (s, 1H), 7.71 (d, 1H), 7.37 (d, 1H), 4.85-4.64 (m, 1H),4.78-4.53 (m, 3H), 4.23-4.17 (m, 4H), 3.62 (appt, 2H), 2.88-2.76 (m,2H), 1.83 (br d, 2H); MS (EI) m/z 442.1 (MH)⁺.

3-(9((1R,4R)-4-hydroxy-1,2,3,4-tetrahydronaphthalen-1-yl)-6-(3-hydroxyazetidin-1-yl)-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.Synthesis of the foregoing purinone is accomplished according to thereaction sequence presented in Route A with the modification that thefirst step involves the reaction of 2,5-diaminio-4,6-dichloropyrimidinewith (1R,4R)-4-amino-1,2,3,4-tetrahydronaphthalen-1-ol) to give(1R,4R)-4-(2,5-diamino-6-chloropyrimidin-4-ylamino)-1,2,3,4-tetrahydronaphthalen-1-ol.A suitable protection for the alcohol is to be used. The subsequentreaction sequence is then carried out as presented to give, afterdeprotection of the alcohol, the desired product.

2-(6-chloro-1H-benzo[d]imidazol-1-yl)-6-(3,3-difluoroazetidin-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.White solid (HCl salt). Yield 57%. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm:10.07 (s, 1H), 8.83 (s, 1H), 7.87 (d, 1H), 7.66 (d, 1H), 4.79 (app t,4H), 4.65-4.60 (m, 1H), 4.23-4.17 (m, 2H), 3.63 (appt, 2H), 2.85-2.78(m, 2H), 1.86 (br d, 2H); MS (EI) m/z 462.1 (MH)⁺.

6-(azetidin-1-yl)-2-(6-chloro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.White solid (TFA salt). ¹H NMR (300 MHz, CDCl₃+CD₃OD) δm, ppm: 9.07 (s,1H), 8.67 (s, 1H), 7.70 (d, 1H), 7.35 (d, 1H), 4.65-4.60 (m, 1H), 4.43(app t, 4H), 4.23-4.17 (m, 2H), 3.62 (appt, 2H), 2.85-2.78 (m, 2H),2.68-2.53 (m, 2H), 1.83 (br d, 2H); MS (EI) m/z 426.1 (MH)⁺.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-6-((S)-3-fluoropyrrolidin-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.White solid. TFA salt. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.20 (br s,1H), 8.40-8.31 (m, 1H), 7.78-7.68 (m, 1H), 7.20-7.15 (m, 1H), 5.46 (d,J_(H-C-F)=52.6 HZ, 1H), 4.67-4.57 (m, 1H), 4.21-3.90 (m, 6H), 3.61(appt, 2H), 2.84-2.76 (m, 2H), 2.48-2.16 (m, 2H), 1.81 (br d, 2H); MS(EI) m/z 442.1 (MH)⁺.

6-(4,4-difluoropiperidin-1-yl)-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.White solid. TFA salt. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.34 (s,1H), 8.31-8.27 (m, 1H), 7.81-7.76 (m, 1H), 7.27-7.20 (m, 1H), 4.67-4.57(m, 1H), 4.20 (dd, 2H), 3.91 (t, 4H), 3.63 (appt, 2H), 2.91-2.76 (m,2H), 2.25-2.11 (m, 4H), 1.84 (br d, 2H); MS (EI) m/z 474.0 (MH)⁺.

6-(cyclobutylamino)-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.Pale yellow solid. TFA salt. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.09(s, 1H), 8.37-8.33 (m, 1H), 7.75-7.69 (m, 1H), 7.17-7.14 (m, 1H),4.73-4.67 (m, 1H), 4.64-4.57 (m, 1H), 4.19 (dd, 2H), 3.62 (appt, 2H),2.87-2.76 (m, 2H), 2.59-2.55 (m, 2H), 2.09-1.81 (m, 6H); MS (EI) m/z424.2 (MH)⁺.

Method 3

Typical procedure for cross-coupling reactions of 6-chloropurinones withorganozinc halides. THF (3 mL) was added to an argon-purged vialcontaining6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(0.2 mmol) and Pd(PPh₃)₄ (0.05 equiv.). The mixture was stirred at roomtemperature for 10 min, and then a 0.5 M solution of an organozincreagent in THF (15 equiv.) was added dropwise at room temperature. Thestirring at room temperature was continued for 15 min, followed bystirring at 50° C. for 3 h. The reaction mixture was left to cool toroom temperature, the solvent was removed in vacuo, acetonitrile wasadded to the residue and the mixture filtered through a Nylon 0.45 μmfilter. Preparative HPLC purification afforded, after evaporation anddrying, the desired compound.

6-cyclobutyl-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.White semi-solid, prepared using a 0.5 M solution of cyclobutylzincbromide in THF. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.26 (s, 1H), 8.49(d, 1H), 7.79-7.73 (m, 1H), 7.22-7.16 (m, 1H), 4.73-4.67 (m, 1H), 4.19(dd, 2H), 3.94-3.82 (m, 1H), 3.63 (appt, 2H), 2.90-2.76 (m, 2H),2.70-2.57 (m, 2H), 2.53-2.42 (m, 2H), 2.31-2.08 (m, 2H), 1.98-1.83 (m,2H); MS (EI) m/z 409.1 (MH)⁺.

Method 4

General procedure for displacement reactions of 6-chloropurinones withalcohols.6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(0.128 mmol) was taken in a dry microwave vial. To it was added analiphatic or aromatic alcohol (1-10 equiv), cesium carbonate (10 equiv.)and DMF. The sealed reaction tube was subjected to microwave heating at200° C. for 4 h. Completion of the reaction was checked by LC-MS.Solvent was removed under high vacuum, the residue was dissolved in MeOHand filtered through a Nylon 0.45 μm filter. Preparative HPLCpurification afforded, after solvent evaporation and drying, the desiredproduct.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-6-pyridin-3-yloxy)-9-(tetrahydro-2H-pyran-4-yl)7H-purin-8(9H)-one

Synthesized using 3-hydroxypyridine (12.2 mg, 0.128 mmol, 1 equiv). ¹HNMR (300 MHz, CD₃OD) δ, ppm: 9.20 (br s, 2H), 8.10 (d, 2H), 7.80 (br m,2H), 7.45 (br m, 1H), 7.15 (br m, 1H), 4.80-4.63 (m, 1H), 4.18 (dd, 2H),3.65 (app t, 2H), 2.90-2.72 (m, 2H), 1.92-1.82 (m, 2H); MS (EI) m/z448.1 (MH)⁺.

6-cyclobutoxy-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one

Synthesized using cyclobutanol (10 μL, 1.28 mmol, 10 equiv.). ¹H NMR(300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.01 (s, 1H), 8.26 (dd, 1H), 7.77 (dd,1H), 7.16 (td, 1H), 5.54-5.44 (m, 1H), 4.68-4.56 (m, 1H), 4.22 (dd, 2H),3.62 (appt, 2H), 2.89-2.72 (m, 2H), 2.70-2.58 (m, 2H), 2.40-2.24 (m,2H), 2.06-1.92 (m, 1H), 1.92-1.80 (m, 3H); MS (EI) m/z 425.1 (MH)⁺.Variant B

Typical Procedure for N-7 alkylation/methylation. To a solution of2-(6-chloro-1H-benzo[d]imidazol-1-yl)-6-(3,3-difluoroazetidin-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(15 mg, 0.033 mmol) in CH₃CN (5 mL) was added polystyrene supported BEMP(2-tert.butylimino-2-diethylamino-1,3-dimethyl-perhydro-1,3,2-diazaphosphorine(Fluka, loading 2.2 mmol/g) (4 equiv.), followed by alkyliodide/iodomethane (6 equiv.). The reaction mixture was stirred at roomtemperature for 1 h. Completion of the reaction mixture was checked byHPLC and MS. The reaction mixture was filtered and the resin was washedwith CH₃CN (5 mL×2) and MeOH (5 mL×2). The washings and the filtratewere combined and concentrated in vacuo to give the desired compound.

2-(6-chloro-1H-benzo[d]imidazol-1-yl)-6-(3,3-difluoroazetidin-1-yl)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.White solid. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.22 (s, 1H), 8.63(s, 1H), 7.73 (br d, 1H), 7.37 (br d, 1H), 4.79 (app t, 4H), 4.65-4.60(m, 1H), 4.23-4.13 (m, 2H), 3.65 (appt, 2H), 3.57 (s, 3H), 2.85-2.78 (m,2H), 1.82 (br d, 2H); MS (EI) m/z 476.2 (MH)⁺.

2-(benzylthio)-6-chloro-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine.To a solution of 2-(benzylthio)-4,6-dichloro-5-nitropyrimidine (WO01/58906 A1) (9 g, 28.5 mmol) in THF (60 mL) and DIEA (9.9 mL, 7.36 g,57 mmol, 2 equiv.) was added drop wise a solution of4-aminotetrahydropyran (2.77 g, 27.4 mmol, 0.96 equiv.) in THF (20 mL)over 15 min. The reaction mixture was stirred at room temperature for 18h. The solvent was then removed in vacuo and the residue purified bycolumn chromatography (silica gel, elution with 4/1 hexanes/ethylacetate) to afford, after evaporation and drying, the desired product asa yellow solid (7.59 g, 70% yield). ¹H NMR (300 MHz, CDCl₃) δ, ppm: 7.86(br d, J=6.8 Hz, 1H), 7.42-7.36 (m, 2H), 7.36-7.27 (m, 3H), 4.38 (s,2H), 4.31-4.20 (m, 1H), 3.96 (dt, J=11.7, 3.5 Hz, 2H), 3.47 (ddd,J=11.7, 11.3, 2.2 Hz, 2H), 1.92 (br dd, J=12.4, 2.2 Hz, 2H), 1.66-1.52(m, 2H).

General procedure for cross-coupling reactions of2-(benzylthio)-6-chloro-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-aminewith organozinc halides. In an oven dried flask under Ar, Pd(PPh₃)₄ (152mg, 0.13 mmol, 0.05 equiv.) was added to a solution of2-(benzylthio)-6-chloro-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine(1 g, 2.63 mmol, 1.0 equiv.) in THF (26 mL). The mixture was stirredunder Ar for 10 min, then it was cooled to 0° C. and the organozinchalide reagent (0.5 M solution in THF, 1.5 equiv.) was added drop wise,under Ar, within 30 min. The reaction mixture was then warmed up to roomtemperature, and subsequently heated to 50° C. for 3 days. Uponcompletion of the reaction, the mixture was poured into a saturatedaqueous NH₄Cl solution. Extraction into ethyl acetate (3×100 mL),washing of the combined organic layers with brine (1×100 mL), drying(Na₂SO₄), and solvent removal in vacuo afforded a brown residue, whichwas purified by column chromatography to give the desired product.

2-(benzylthio)-6-(2-chlorophenyl)-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine.Yellow solid. 0.81 g, 68% yield. Synthesized using 2-chlorophenylzinciodide, 0.5 M in THF (7.9 mL, 3.95 mmol, 1.5 equiv.). Columnchromatography purification: silica gel, elution with 6/1 hexanes/ethylacetate. ¹H NMR (300 MHz, CDCl₃) δ, ppm: 8.20 (br d, 1H), 7.42-7.27 (m,9H), 4.41 (s, 2H), 4.41-4.32 (m, 1H, overlapping with 4.41 ppm),4.02-3.94 (m, 2H), 3.55-3.47 (m, 2H), 1.99-1.94 (m, 2H), 1.64-1.56 (m,2H); MS (EI) m/z 457.1 (MH)⁺.

4-(2-(benzylthio)-5-nitro-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidin-4-yl)benzonitrile.0.39 g, 33% yield. Synthesized using 4-cyanophenylzinc bromide, 0.5 M inTHF (8 mL, 4 mmol, 1.5 equiv.). Column chromatography purification:silica gel, gradual elution with 9/1 to 4/1 hexanes/ethyl acetate. ¹HNMR (300 MHz, CDCl₃) δ, ppm: 7.88 (br d, 1H), 7.73 (d, 2H), 7.54 (d,2H), 7.41-7.27 (m, 5H), 4.41 (s, 2H), 4.41-4.32 (m, 1H, overlapping with4.41 ppm), 4.02-3.96 (m, 2H), 3.55-3.46 (m, 2H), 1.99-1.94 (m, 2H),1.64-1.55 (m, 2H); MS (EI) m/z 448.1 (MH)⁺.

General procedure for the oxidation of2-(benzylthio)-6-substituted-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amineto2-(benzylsulfonyl)-6-substituted-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine.A solution of mCPBA (4.11 mmol, 2.3 equiv.) in methylene chloride (20mL) was dried over anhydrous Na₂SO₄ and added dropwise, under Ar, to asolution of2-(benzylthio)-6-substituted-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine(1.78 mmol, 1.0 equiv.) in 50 mL of anhydrous methylene chloride at 0°C. The reaction mixture was then stirred at room temperature for 18 h.Upon completion of the reaction, the mixture was washed repeatedly witha saturated aqueous solution of NaHCO₃ (6×100 mL), the organic layer waswashed with water (1×100 mL), dried (Na₂SO₄), and the solvent removed invacuo.

2-(benzylsulfonyl)-6-(2-chlorophenyl)-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine.Orange solid. 0.87 g, yield quantitative. Synthesized from2-(benzylthio)-6-(2-chlorophenyl)-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine.¹H NMR (300 MHz, CDCl₃) δ, ppm: 7.92 (br d, 1H), 7.50-7.27 (m, 9H), 4.74(s, 2H), 4.51-4.41 (m, 1H), 4.02-3.98 (m, 2H), 3.61-3.52 (m, 2H),2.07-2.01 (m, 2H), 1.71-1.57 (m, 2H); MS (EI) m/z 489.2 (MH)⁺.

4-(2-(benzylsulfonyl)-5-nitro-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidin-4-yl)benzonitrile.Yield quantitative. Synthesized from4-(2-(benzylthio)-5-nitro-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidin-4-yl)benzonitrile.¹H NMR (300 MHz, CDCl₃) δ, ppm: 7.96 (br d, 1H), 7.78 (d, 2H), 7.62 (d,2H), 7.47-7.28 (m, 5H), 4.75 (s, 2H), 4.51-4.41 (m, 1H), 4.04-3.98 (m,2H), 3.61-3.55 (m, 2H), 2.04-2.00 (m, 2H), 1.71-1.57 (m, 2H); MS (EI)m/z 480.2 (MH)⁺.

General procedure for the displacement of2-(benzylsulfonyl)-6-substituted-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-aminewith ammonia. To a solution of2-(benzylsulfonyl)-6-substituted-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine(1.58 mmol) in 80 mL anhydrous methylene chloride at 0° C. was addeddropwise 4 mL of a 7 M solution of ammonia in methanol. Upon completionof the reaction (5-10 min), the solvent was removed in vacuo. Theresidue was taken in ethyl acetate/ethyl ether 100 mL/100 mL, washedwith a saturated aqueous solution of ammonium chloride (2×100 mL),followed by a saturated aqueous solution of sodium carbonate (2×100 mL),and brine (1×100 mL). The organic layer was dried (Na₂SO₄), thenconcentrated in vacuo.

6-(2-chlorophenyl)-5-nitro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4-diamine.Pale yellow solid. 0.48 g, 87% yield. Synthesized from2-(benzylsulfonyl)-6-(2-chlorophenyl)-5-nitro-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine.¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 7.52-7.29 (m, 4H), 4.51-4.41 (m,1H), 4.04-3.90 (m, 2H), 3.78-3.52 (m, 2H), 2.07-2.01 (m, 2H), 1.71-1.64(m, 2H); MS (EI) m/z 350.3 (MH)⁺.

4-(2-amino-5-nitro-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidin-4-yl)benzonitrile.Pale yellow solid. 75% yield. Synthesized from4-(2-(benzylsulfonyl)-5-nitro-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidin-4-yl)benzonitrile.MS (EI) m/z 341.2 (MH)⁺.

General procedure for the reduction of6-substituted-5-nitro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4-diamine.To6-substituted-5-nitro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4-diamine(1.57 mmol, 1.0 equiv.) was added ethyl acetate (75 mL), followed by Ptcatalyst, 0.5% sulfur, 5% on activated carbon (Alfa Aesar) (304 mg,0.078 mmol, 0.05 equiv.) with stirring at room temperature underhydrogen (35 psi) for 6 h (HPLC monitoring). Filtration of the reactionmixture over a small plug of celite, thorough washing with ethyl acetateand methanol, evaporation of solvent and column chromatography of theresidue afforded the desired product.

6-(2-chlorophenyl)-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine.Yellow solid. 0.26 g, 63% yield based on 84% conversion. Synthesizedfrom6-(2-chlorophenyl)-5-nitro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4-diamine.Column chromatography on silica gel, elution with 9/1 methylenechloride/ethyl acetate to 4/1 methylene chloride/ethyl acetate with upto 10% methanol. ¹H NMR (300 MHz, CD₃OD) δ, ppm: 7.52-7.42 (m, 1H),7.35-7.27 (m, 3H), 4.21-4.10 (m, 1H), 3.93-3.77 (m, 2H), 3.51-3.42 (m,2H), 1.97-1.91 (m, 2H), 1.55-1.50 (m, 2H); MS (EI) m/z 320.2 (MH)⁺.

General procedure for the selective formation of the purinone from6-substituted-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine. Asolution of6-substituted-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine(0.5 mmol, 1.0 equiv.) in 90 mL of anhydrous THF was added dropwise,over 30 min, to a solution of phosgene in THF (0.5 mL of a 20% solutionof phosgene in toluene, 0.95 mmol, 1.9 equiv., diluted with 10 mL ofanhydrous THF) at −78° C. under Ar. The reaction mixture was left togradually warm up to room temperature, then stirred at room temperaturefor 30 min. It was purged with air for 30 min. The solvent was thenremoved in vacuo, and the residue was taken in ethyl acetate/saturatedaqueous NaHCO₃. The layers were separated, and the aqueous layerextracted with ethyl acetate three times. The combined organic extractswere put together, dried (anhydrous Na₂SO₄) and the solvent removed invacuo. Chromatographic purification afforded the desired product.

2-amino-6-(2-chlorophenyl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.White solid. 150 mg, 87% yield. Synthesized from6-(2-chlorophenyl)-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine.Preparative TLC purification on silica gel, elution with 3% methanol inmethylene chloride. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 7.57-7.41 (m,4H), 4.58-4.47 (m, 1H), 4.15 (ddd, 2H), 3.58 (appt, J=11.7 Hz, 2H), 2.82(ddd, 2H), 1.74 (br d, J=10.6 Hz, 2H); MS (EI) m/z 346.3 (MH)⁺.

4-(2-amino-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-6-yl)benzonitrile.Synthesized from4-(2,5-diamino-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidin-4-yl)benzonitrile.¹H NMR (300 MHz, CDCl₃) δ, ppm: 7.98 (d, 2H), 7.82 (d, 2H), 7.42 (br d,1H), 4.58-4.47 (m, 1H), 4.18-4.09 (m, 2H), 3.56 (app t, 2H), 2.93 (br s,2H), 2.88-2.70 (m, 2H), 1.78-1.67 (m, 2H); MS (EI) m/z 337.3 (MH)⁺.

General procedure for the Buchwald-Hartwig Palladium-catalyzed C—N crosscoupling. An oven-dried vial was charged with2-amino-6-substituted-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(0.29 mmol, 1 equiv.), freshly grounded cesium carbonate (132 mg, 0.41mmol, 1.4 equiv.), Pd(OAc)₂(20 mg, 0.003 mmol, 0.1 equiv.), racemicBINAP (27 mg, 0.04 mmol, 0.15 equiv.) and1-bromo-5-fluoro-2-nitrobenzene (83 mg, 0.38 mmol, 1.3 equiv.), followedby anhydrous toluene (2 mL). The vial was purged with Ar for 3 min, thenclosed and heated at 80° C. for 3 days. The reaction mixture was allowedto stand overnight at room temperature and poured into saturated aqueousNH₄Cl (10 mL). To this mixture, saturated aqueous Na₂EDTA (10 mL) wasadded and the mixture was stirred for 10 min. The mixture was extractedwith ethyl acetate (3×20 mL), the collected organic layers were washedwith brine, dried (anhydrous Na₂SO₄) and the solvent evaporated invacuo. Column chromatography of the residue on silica gel, elution with1% methanol in 4/2/1 hexane/methylene chloride/ethyl acetate afforded,after evaporation and drying, the desired product.

6-(2-chlorophenyl)-2-(5-fluoro-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.Yellow solid. 22 mg, 16% yield. Synthesized from2-amino-6-(2-chlorophenyl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.¹H NMR (300 MHz, CDCl₃) δ, ppm: 10.77 (s, 1H), 8.98 (dd, 1H), 8.34 (dd,1H), 7.99 (s, 1H), 7.69-7.65 (m, 1H), 7.60-7.48 (m, 3H), 6.75-6.68 (m,1H), 4.66-4.57 (m, 1H), 4.19 (dd, 2H), 3.59 (app t, J=11.7 Hz, 2H), 2.81(ddd, 2H), 1.81 (br d, J=10.3 Hz, 2H); MS (EI) m/z 485.3 (MH)⁺.

4-(2-(5-fluoro-2-nitrophenylamino)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-6-yl)benzonitrile.Yellow solid. 15% yield. Synthesized from4-(2-amino-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-6-yl)benzonitrile.MS (EI) m/z 476.2 (MH)⁺.

General procedure for the reduction of the nitrophenyl group in6-substituted-2-(5-fluoro-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-onewith iron powder. To a vial containing6-substituted-2-(5-fluoro-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(0.045 mmol) was added glacial acetic acid (1 mL), water (2.5 mL) andethanol (5 mL), followed by iron powder (26 mg, 0.45 mmol, 10 equiv.)and the resulting mixture heated at 90° C. for 15 min (HPLC monitoring).The reaction mixture was left to cool down to room temperature,concentrated ammonium hydroxide solution was added to bring the pH tobasic, and the mixture was stirred for 10 min. The aqueous layer dilutedwith water (10 mL) was extracted with ethyl acetate (3×20 mL), thecombined organic layers were washed with brine, dried (anhydrousNa₂SO₄), and the solvent removed in vacuo to give the desired product.This material was used in the next step without further purification.

2-(2-amino-5-fluorophenylamino)-6-(2-chlorophenyl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.Pale yellow solid. 21 mg, yield quantitative. Synthesized from6-(2-chlorophenyl)-2-(5-fluoro-2-nitrophenylamino)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.MS (EI) m/z 455.3 (MH)⁺.

4-(2-(2-amino-5-fluorophenylamino)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-6-yl)benzonitrile.Yield quantitative. Synthesized from4-(2-(5-fluoro-2-nitrophenylamino)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-6-yl)benzonitrile.MS (EI) m/z 446.3 (MH)⁺.

General procedure for closing the benzimidazole ring. To a vialcontaining crude2-(2-amino-5-fluorophenylamino)-6-substituted-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(0.045 mmol) was added anhydrous methanol (4 mL), followed by anhydroustrimethylorthoformate (0.1 mL) and methane sulfonic acid (catalyticamount) and the reaction mixture was stirred under Ar at roomtemperature for 3 h (HPLC monitoring). Preparative TLC purification(silica gel, elution with 2.5% methanol in methylene chloride) afforded,after evaporation of solvent and drying, the desired product.

6-(2-chlorophenyl)-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.White solid. 12 mg, 57% yield on two steps. Synthesized from2-(2-amino-5-fluorophenylamino)-6-(2-chlorophenyl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.¹H NMR (300 MHz, CDCl₃) δ, ppm: 9.12 (s, 1H), 8.32 (br d, 1H), 7.82-7.71(m, 3H), 7.62-7.51 (m, 3H), 7.16-7.09 (m, 1H), 4.72-4.64 (m, 1H),4.24-4.21 (m, 2H), 3.61 (app t, J=11.7 Hz, 2H), 2.87-2.79 (m, 2H), 1.88(br d, J=11.4 Hz, 2H); MS (EI) m/z 465.2 (MH)⁺.

4-(2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-6-yl)benzonitrile.11% yield on two steps. Synthesized from4-(2-(2-amino-5-fluorophenylamino)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-6-yl)benzonitrile.¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.12 (s, 1H), 8.32 (br d, 1H),8.18 (d, 2H), 7.93 (d, 2H), 7.85-7.72 (m, 1H), 7.22-7.13 (m, 1H),4.68-4.62 (m, 1H), 4.27-4.16 (m, 2H), 3.62 (app t, 2H), 2.92-2.77 (m,2H), 1.86 (br dd, 2H); MS (EI) m/z 456.2 (MH)⁺.

2-amino-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.Anhydrous ethanol (2 mL) and anhydrous 1,4-dioxane (2 mL) were added toan argon-purged vial containing2-amino-6-chloro-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one (seepreparation Route A, 100 mg, 0.37 mmol), pyridine-4-boronic acid (69 mg,0.56 mmol, 1.5 equiv.), K₃PO₄ (157 mg, 0.74 mmol, 2.0 equiv.), Pd(OAc)₂(12.5 mg, 0.018 mmol, 0.05 equiv.) and D-tBPF1,1′-bis(di-tbutylphosphino)ferrocene (8.8 mg, 0.018 mmol, 0.05 equiv.)and the mixture was heated for 2.5 h in the microwave oven at 150° C.After cooling to ambient temperature, the reaction mixture was dilutedwith methanol (20 mL), filtered through a Nylon 0.45 μm filter and thefiltrate concentrated in vacuo. Preparative HPLC purification of theresidue afforded, after evaporation and drying, the desired compound asa yellow solid (59.2 mg, 51% yield) TFA salt. ¹H NMR (300 MHz, CD₃OD) δ,ppm: 8.92 (d, J=6.4 Hz, 2H), 8.17 (dd, J=5.1, 1.5 Hz, 2H), 4.87-4.55 (m,1H), 4.10 (dd, J=11.4, 4.3 Hz, 2H), 3.56 (ddd, J=11.3, 6.5, 4.7 Hz, 2H),2.74 (tdd, J=12.6, 12.5, 4.6 Hz, 2H), 1.77 (dd, J=12.4, 2.4 Hz, 2H); MS(EI) m/z 313.3 (MH)⁺.

2-(5-chloro-2-nitrophenylamino)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.To an oven-dried vial was added2-amino-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(68 mg, 0.22 mmol, 1 equiv.) in anhydrous toluene (1 mL), then freshlygrounded cesium carbonate (99.4 mg, 0.31 mmol, 1.4 equiv.) with stirringat room temperature under Ar. After 20 min, Pd(OAc)₂ (15 mg, 0.02 mmol,0.1 equiv.), racemic BINAP (20.5 mg, 0.03 mmol, 0.15 equiv.) and1-bromo-5-chloro-2-nitrobenzene (WO 02/053545 A1) (67.6 mg, 0.29 mmol,1.3 equiv.) were added as solids, followed by anhydrous DMSO (1 mL). Thevial was purged with Ar for 3 min, then closed and heated in a microwaveoven at 170° C. for 45 min. The reaction mixture was cooled to roomtemperature, diluted with ethyl acetate/methylene chloride, filteredthrough a Nylon 0.45 μm filter, and the filtrate was concentrated invacuo. The resulting residue was purified using preparative TLC (silicagel, 7.5% methanol in methylene chloride) to give the desired product asa brown solid (11 mg, 11% yield). ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm:9.13 (s, 1H), 8.63 (br m, 2H), 8.13 (d, J=8.9 Hz, 1H), 7.83 (br m, 2H),6.91 (d, J=8.6 Hz, 1H), 4.70-4.50 (m, 1H), 4.10-4.03 (m, 2H), 3.51-3.43(m, 2H), 2.68-2.65 (m, 2H), 1.71-1.66 (m, 2H); MS (EI) m/z 468.4 (MH)⁺.

2-(2-amino-5-chlorophenylamino)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.To a vial containing2-(5-chloro-2-nitrophenylamino)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(7 mg, 0.015 mmol) was added glacial acetic acid (0.5 mL), water (1.25mL) and ethanol (2.5 mL), followed by iron powder (8.4 mg, 0.15 mmol, 10equiv.) and the resulting mixture heated at 90° C. for 15 min (HPLCmonitoring). The reaction mixture was left to cool down to roomtemperature, concentrated ammonium hydroxide solution was added to bringthe pH to basic, and the mixture was stirred for 10 min. The aqueouslayer was extracted with ethyl acetate, the combined organic layers werewashed with brine, dried (anhydrous Na₂SO₄), and the solvent removed invacuo to give the desired product as a pale yellow solid (yieldquantitative). This material was used in the next step without furtherpurification. MS (EI) m/z 438.3 (MH)⁺.

2-(6-chloro-1H-benzo[d]imidazol-1-yl)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.To a vial containing crude2-(2-amino-5-chlorophenylamino)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(0.015 mmol) was added anhydrous methanol (2.5 mL), followed byanhydrous trimethylorthoformate (0.5 mL) and p-toluene sulfonic acid (1mg) and the reaction mixture was stirred under Ar at room temperaturefor 4 h (HPLC monitoring). Preparative HPLC purification, afterevaporation of solvent and drying, provided the desired product (1.2 mg,18% yield on two steps), as a yellow solid. TFA salt. ¹H NMR (300 MHz,CD₃OD) δ, ppm: 9.18 (s, 1H), 8.83 (br m, 2H), 8.71 (s, 1H), 8.07 (br m,2H), 7.76 (d, J=8.6 Hz, 1H), 7.41 (d, J=8.9 Hz, 1H), 4.76-4.72 (m, 1H),4.26-4.23 (m, 2H), 3.66 (app t, J=11.5 Hz, 2H), 2.90-2.87 (m, 2H), 1.91(br d, J=12.8 Hz, 2H); MS (EI) m/z 448.3 (MH)⁺.

Modification of the General Procedure to allow access to6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-2-(6-(trifluoromethoxy)-1H-benzo[d]imidazol-1-yl)-7H-purin-8(9H)-one

2-(2-(2,5-dimethyl-1H-pyrrol-1-yl)-5-(trifluoromethoxy)phenylamino)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.Pale yellow solid. Synthesized following the general procedure for theBuchwald-Hartwig Palladium-catalyzed C—N cross coupling using2-amino-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(86 mg, 0.27 mmol) and commercially available1-[2-bromo-4-(trifluoromethoxy)phenyl-2,5-dimethyl-1H-pyrrole](Maybridge, 1.3 equiv.) as the aryl bromide. 11 mg (12% yield based on58% conversion). MS (EI) m/z 566.4 (MH)⁺.

2-(2-amino-5-(trifluoromethoxy)phenylamino)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.2-(2-(2,5-dimethyl-1H-pyrrol-1-yl)-5-(trifluoromethoxy)phenylamino)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one(0.018 mmol) was reacted with hydroxylamine hydrochloride (5 equiv.) ina mixture of ethanol and water (1.6 mL/0.6 mL) at reflux for 20 h. Uponcompletion of the reaction (monitoring by HPLC and MS), the solvent wasremoved in vacuo, the material thoroughly dried and used in the nextstep without further purification. MS (EI) m/z 488.4 (MH)⁺.

6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-2-(6-(trifluoromethoxy)-1H-benzo[d]imidazol-1-yl)-7H-purin-8(9H)-one.Yellow oil. TFA salt. Synthesized from2-(2-amino-5-(trifluoromethoxy)phenylamino)-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-onefollowing the general procedure for closing the benzimidazole ring. ¹HNMR (300 MHz, CD₃OD+CDCl₃) δ, ppm: 9.30 (br s, 1H), 9.00-8.85 (br m,1H), 8.67 (s, 1H), 8.20-8.10 (m, 2H), 7.84-7.67 (m, 1H), 7.40-7.22 (m,2H), 4.76-4.72 (m, 1H), 4.26-4.13 (m, 2H), 3.62 (app t, 2H), 2.90-2.80(m, 2H), 1.90 (br d, 2H); MS (EI) m/z 498.3 (MH)⁺.

6-chloro-8-methyl-9-(tetrahydro-2H-pyran-4-yl)-9H-purin-2-amine.N,N-Dimethylacetamide (8 mL) containing 0.2% (v) glacial acetic acid wasadded to 6-chloro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine(preparation Route A) (400 mg, 1.64 mmol), followed bytriethylorthoacetate (0.8 mL, 5.32 mmol, 3.2 equiv.), and the reactionmixture was heated at 160° C. in a closed vial for two days. The mixturewas then poured into water, and the pH was brought to basic using asaturated solution of sodium bicarbonate. Multiple extractions withmethylene chloride, washing of the combined organic layers with water,and evaporation of the solvent in vacuo gave a dark brown residue, whichwas purified by column chromatography (silica gel, gradual elution with2% methanol in 4/2/1 hexanes/methylene chloride/ethyl acetate to 3%methanol in 4/2/1 hexanes/methylene chloride/ethyl acetate) to give 330mg (75% yield) of desired product as a pale yellow solid. ¹H NMR (300MHz, CD₃OD) δ, ppm: 4.67-4.57 (m, 1H), 4.11 (dd, J=11.5, 4.6 Hz, 2H),3.61 (td, J=12.0, 1.5 Hz, 2H), 2.91-2.76 (m, 2H), 2.72 (s, 3H), 1.86 (brdd, J=12.5, 2.4 Hz, 2H); MS (EI) m/z 268.1 (MH)⁺.

In an alternative procedure, the reaction was carried out in parallelusing 10 vials. To each vial was added6-chloro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine(preparation Route A) (400 mg, 1.64 mmol), N,N-dimethyl acetamide (8mL), glacial acetic acid (16 μL, 0.2% vol) and triethylorthoacetate (0.8mL, 5.32 mmol) and the mixture was stirred at 160° C. After 3 days, thereaction was complete (by LC-MS) and the reaction mixture, containingdesired product MS (EI) m/z 268.1 (MH)⁺, as well as acetylated productMS (EI) m/z 310.2 (MH)⁺, was cooled to room temperature. To each vialwas then added 1 mL of a 1 M aqueous solution of HCl, with stirring at100° C. After 1 h at 100° C., LC-MS analysis indicated only desiredproduct, in a clean reaction. Upon cooling to room temperature, a whiteprecipitate formed. Filtration under vacuum, followed by washing of theprecipitate with portions of methanol provided the desired product(combined yield 2.98 g for 10 vials, 68% yield) as a white solid in pureform. ¹H NMR (300 MHz, d₆-DMSO) δ, ppm: 6.76 (s, 2H), 4.41 (m, 1H), 3.97(m, 2H), 3.45 (m, 2H), 3.33 (s, 3H), 2.63 (m, 2H), 1.74 (m, 2H); MS (EI)m/z 268.1 (MH)⁺.

8-methyl-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-9H-purin-2-amine.Anhydrous ethanol (4 mL) was added to an argon-purged vial containing6-chloro-8-methyl-9-(tetrahydro-2H-pyran-4-yl)-9H-purin-2-amine (140 mg,0.52 mmol), pyridine-4-yl-boronic acid (97 mg, 0.79 mmol, 1.5 equiv.),K₂CO₃ (109 mg, 0.79 mmol, 1.5 equiv.), and Pd(PPh₃)₄ (30 mg, 0.026 mmol,0.05 equiv.) and the mixture was heated for 30 min in the microwave ovenat 150° C. After cooling to ambient temperature, the reaction mixturewas diluted with methanol (8 mL), filtered through a Nylon 0.45 μmfilter and the filtrate concentrated in vacuo. Preparative HPLCpurification of the residue afforded, after evaporation and drying, thedesired compound as an orange solid (160 mg, 99% yield). TFA salt. ¹HNMR (300 MHz, CD₃OD) δ, ppm: 9.09 (d, J=6.6 Hz, 2H), 8.95 (d, J=6.2 Hz,2H), 4.61-4.52 (m, 1H), 4.11 (dd, J=11.5, 4.5 Hz, 2H), 3.61 (app t,J=11.3 Hz, 2H), 2.96-2.82 (m, 2H), 2.68 (s, 3H), 1.84 (dd, J=12.7, 2.3Hz, 2H); MS (EI) m/z 311.3 (MH)⁺.

N-(5-chloro-2-nitrophenyl)-8-methyl-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-9H-purin-2-amine.To an oven-dried vial was added8-methyl-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-9H-purin-2-amine(56 mg, 0.18 mmol, 1 equiv.) in anhydrous toluene (1 mL), then freshlygrounded cesium carbonate (82 mg, 0.25 mmol, 1.4 equiv.) with stirringat room temperature under Ar. After 10 min, Pd(OAc)₂ (13.5 mg, 0.02mmol, 0.1 equiv.), racemic BINAP (17 mg, 0.03 mmol, 0.15 equiv.) and1-bromo-5-chloro-2-nitrobenzene (WO 02/053545 A1) (55 mg, 0.23 mmol, 1.3equiv.) were added as solids. The vial was purged with Ar for 3 min,then closed and heated in a microwave oven at 170° C. for 2 h. Thereaction mixture was cooled to room temperature, diluted with ethylacetate/methylene chloride, filtered through a Nylon 0.45 μm filter, andthe filtrate was concentrated in vacuo. The resulting residue waspurified using preparative TLC (silica gel, 7.5% methanol in methylenechloride) to give the desired product as a yellow solid (18 mg, 21%yield). ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.30 (s, 1H), 8.77 (br m,2H), 8.64 (br m, 2H), 8.26 (d, J=8.9 Hz, H), 7.05 (d, J=7.9 Hz, 1H),4.60-4.56 (m, 1H), 4.26-4.22 (m, 2H), 3.64 (app t, J=11.9 Hz, 2H),2.94-2.90 (m, 2H), 2.76 (s, 3H), 1.91 (br d, J=11.2 Hz, 2H); MS (EI) m/z466.3 (MH)⁺.

5-chloro-N¹-(8-methyl-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-9H-purin-2-yl)benzene-1,2-diamine.To a vial containingN-(5-chloro-2-nitrophenyl)-8-methyl-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-9H-purin-2-amine(16.3 mg, 0.035 mmol) was added glacial acetic acid (1 mL), water (2.5mL) and ethanol (5 mL), followed by iron powder (20 mg, 0.35 mmol, 10equiv.) and the resulting mixture heated at 90° C. for 15 min (HPLCmonitoring). The reaction mixture was left to cool down to roomtemperature, concentrated ammonium hydroxide solution was added to bringthe pH to basic, and the mixture was stirred for 10 min. The aqueouslayer was extracted with ethyl acetate, the combined organic layers werewashed with brine, dried (anhydrous Na₂SO₄), and the solvent removed invacuo to give the desired product as a pale yellow solid (yieldquantitative). This material was used in the next step without furtherpurification. MS (EI) m/z 436.4 (MH)⁺.

2-(6-chloro-1H-benzo[d]imidazol-1-yl)-8-methyl-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-9H-purine.To a vial containing crude5-chloro-N¹-(8-methyl-6-(pyridin-4-yl)-9-(tetrahydro-2H-pyran-4-yl)-9H-purin-2-yl)benzene-1,2-diamine(0.035 mmol) was added anhydrous methanol (2.5 mL), followed byanhydrous trimethylorthoformate (0.5 mL) and methane sulfonic acid(catalytic amount) and the reaction mixture was stirred under Ar at roomtemperature for 4 h (HPLC monitoring). Preparative HPLC purification,after evaporation of solvent and drying, provided the desired product(3.5 mg, 22% yield on two steps), as a yellow oil. TFA salt. ¹H NMR (300MHz, CD₃OD+CDCl₃) δ, ppm: 9.32 (s, 1H), 8.91 (br m, 4H), 8.79 (s, 1H),7.78 (d, J=8.6 Hz, 1H), 7.43 (d, J=8.6 Hz, 1H), 4.76-4.72 (m, 1H),4.32-4.29 (m, 2H), 3.72 (app t, J=11.7 Hz, 2H), 3.08-3.05 (m, 2H), 2.87(s, 3H), 2.02 (br d, J=10.9 Hz, 2H); MS (EI) m/z 446.3 (MH)⁺.

Trans-4-(2-(6-chloro-1H-benzo[d]imidazol-1-yl)-8-methyl-6-(pyridine-4-yl)-9H-purin-9-yl)cyclohexanol.Synthesis of the foregoing purine is accomplished according to thereaction sequence presented in Route D with the modification that thefirst step involves the reaction oftrans-4-(2,5-diamino-6-chloropyrimidin-4-ylamino)cyclohexanol instead of6-chloro-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-2,4,5-triamine.Trans-4-(2,5-diamino-6-chloropyrimidin-4-ylamino)cyclohexanol isprepared as described in Route J.

(R)-6-chloro-N⁴-(8-fluorochroman-4-yl)pyrimidine-2,4,5-triamine.2,5-diamino-4,6-dichloropyrimidine (1.5 g, 8.37 mmol),(R)-8-fluorochroman-4-amine hydrochloride (see preparation below) (1.7g, 8.37 mmol, 1 equiv.), sodium bicarbonate (2.46 g, 29.3 mmol, 3.5equiv.) and 1-butanol (30 mL) were heated together at 150° C. in asealed tube. After 3 days, when the reaction appeared to be complete (byHPLC), the reaction mixture was cooled to room temperature and thesolvent was removed in vacuo. The residue was purified by flashchromatography (silica gel, elution with 1/1 ethyl acetate/methylenechloride) to give 2.01 g (78% yield) of the desired product as a yellowsolid. ¹H NMR (300 MHz, CD₃OD) δ, ppm: 6.97-6.86 (m, 2H), 6.77-6.72 (m,1H), 5.36 (m, 1H), 4.25 (m, 2H), 2.10 (m, 2H); MS (EI) m/z 310.1 (MH)⁺.

Synthesis and optical resolution of 8-fluorochroman-4-amine wasperformed as described in WO 2006/108103 A1

3-(2-fluorophenoxy)propanoic acid (2). A mixture of 2-fluorophenol (1)(15 g), 3-bromopropanoic acid (20 g) and NaOH (11 g) was refluxed in 50mL of water. The solution was cooled to room temperature and acidifiedto pH 2 with 3 M HCl. The resulting precipitate was isolated byfiltration to yield 9.27 g of title compound as a white solid. Thefiltrate was extracted three times with EtOAc to yield 2.5 g of lesspure compound (2).

8-fluorochroman-4-one (3). Oxalyl chloride (8.79 mL) and one drop of DMFwere added to an ice cold solution of 3-(2-fluorophenoxy)propanoic acid(9.27 g) in methylene chloride (50 mL). The solution was stirred at 0°C. for two hours, aluminum chloride (7.39 g, 55.42 mM) was added and thesolution was stirred for 16 hours at room temperature. The mixture waspoured onto ice water, and extracted three times with methylenechloride. The combined organics were washed with 0.5M NaOH and brine,dried, evaporated, and purified by column chromatography (eluting with20% EtOAc/Hex) to give 8-fluorochroman-4-one (3) (8.20 g, 98%).

8-fluorochroman-4-amine (5). A round bottom flask was charged with8-fluorochroman-4-one (8.2 g), hydroxylamine hydrochloride (3.78 g) andsodium acetate (4.46 g). A reflux condenser was added, the flask waspurged with argon, dry EtOH (20 mL) was added, and the mixture wasstirred at reflux for 18 hours. The solution was cooled to roomtemperature, diluted with EtOAc, and washed with water. The organicphase was dried, and evaporated to give the intermediate8-fluorochroman-4-one oxime (4), which was reduced with Raney Nickel inEtOH at 50 PSI to yield the title amine (5) (4.69 g, 57%).

Resolution of 8-fluorochroman-4-amine (WO 2006/108103)

A mixture of 8-fluorochroman-4-amine (3.40 g), methyl 2-methoxyacetate(2.44 g) and Novozyme 435 (Aldrich, 0.68 g) in anhydrous tert-butylmethyl ether (75 mL) was heated at reflux under argon for two hours (atwhich time the ratio of acylated to unacylated product was 1:1 by HPLC).The solid that formed upon cooling was collected via filtration anddissolved in EtOAc. The mixture was filtered to remove the biocatalystand washed once with 0.5M HCl to remove any lingering (S)-amine. Thesolvent was evaporated and the product was recrystallized fromtert-butyl methyl ether to yield(R)—N-(8-fluorochroman-4-yl)-2-methoxyacetamide (0.78 g). The reactionsolvent and recrystallization mother liquor was washed three times with0.5 M HCl and concentrated to yield additional(R)-N-(8-fluorochroman-4-yl)-2-methoxyacetamide (0.83 g). The combinedacidic aqueous layers were made basic by NaOH and extracted withmethylene chloride to yield (S)-8-fluorochroman-4-amine (5a) (1.6 g). Asolution of (R)-N-(8-fluorochroman-4-yl)-2-methoxyacetamide (0.78 g) in8M HCl in EtOH (50 mL) was heated at reflux for four hours. The solventswere removed from the cooled reaction mixture, the resulting solid wastaken up in 50 mL of 0.5M NaOH, salted out with NaCl_((s)), andextracted four times with methylene chloride to yield(R)-8-fluorochroman-4-amine (0.48 g (87%)) (5b). The % ee was checkedvia chiral HPLC: Chiralcel OD-H (0.46×25 cm analytical column, DaicelChemical Industries) method: isocratic 5% (0.05% TFA/EtOH) 95% (0.05%TFA/Hex), Rt=7.2 min (S)-enantiomer, Rt=9.2 min (R)-enantiomer.

(R)-2-amino-6-chloro-9-(8-fluorochroman-4-yl)-7H-purin-8(9H)-one. To asolution of(R)-6-chloro-N⁴-(8-fluorochroman-4-yl)pyrimidine-2,4,5-triamine (0.9 g,2.91 mmol) in 900 mL of anhydrous THF at −78° C. under Ar was addeddropwise, over 20 min, a solution of phosgene in THF (2.6 mL of a 20%solution of phosgene in toluene, 4.94 mmol, 1.7 equiv., diluted with 26mL of anhydrous THF). The reaction mixture was left to gradually warm upto room temperature over 16 h. It was purged with air for 30 min. Thesolvent was then removed in vacuo, and the residue was taken in ethylacetate/saturated aqueous NaHCO₃. The layers were separated, and theaqueous layer extracted with ethyl acetate three times. The combinedorganic extracts were put together, dried (anhydrous Na₂SO₄) and thesolvent removed in vacuo. The residue was purified by flashchromatography (silica gel, gradual elution with 6/1 to 4/1 methylenechloride/ethyl acetate) to give 0.78 g (80% yield) of the desiredproduct as a pale yellow solid. ¹H NMR (300 MHz, CD₃OD) δ, ppm:7.13-7.06 (m, 1H), 6.90-6.82 (m, 1H), 6.75-6.72 (m, 1H), 5.92-5.82 (m,1H), 4.80-4.73 (m, 1H), 4.50-4.42 (m, 1H), 3.20-3.05 (m, 1H), 2.42-2.30(m, 1H); MS (EI) m/z 336.3 (MH)⁺.

(R)-6-chloro-2-(5-fluoro-2-nitrophenylamino)-9-(8-fluorochroman-4-yl)-7H-purin-8(9H)-one.To an oven-dried vial was added(R)-2-amino-6-chloro-9-(8-fluorochroman-4-yl)-7H-purin-8(9H)-one (152mg, 0.45 mmol, 1 equiv.) in anhydrous toluene (2.5 mL), followed byfreshly grounded cesium carbonate (207 mg, 0.64 mmol, 1.4 equiv.) withstirring at room temperature under Ar. After 10 min, Pd(OAc)₂ (30.3 mg,0.045 mmol, 0.1 equiv.), racemic BINAP (42 mg, 0.067 mmol, 0.15 equiv.)and 1-bromo-5-fluoro-2-nitrobenzene (130 mg, 0.59 mmol, 1.3 equiv.) wereadded as solids. The vial was purged with Ar for 3 min, then closed andheated at 80° C. for three days. The reaction mixture was cooled to roomtemperature, then poured into saturated aqueous NH₄Cl (10 mL). To thismixture, saturated aqueous Na₂EDTA (10 mL) was added and the mixture wasstirred for 10 min. Then the reaction mixture was extracted with ethylacetate (4×20 mL), the collected organic layers were washed with brine,dried (anhydrous Na₂SO₄) and the solvent evaporated in vacuo. Columnchromatography of the residue on silica gel (gradual elution with 1%methanol in 4/2/1 hexane/methylene chloride/ethyl acetate to 2% methanolin 4/2/1 hexane/methylene chloride/ethyl acetate) afforded, afterevaporation and drying, the desired product as an orange solid (76 mg,46% yield based on 78% conversion), along with 33 mg of recoveredstarting material. ¹H NMR (300 MHz, CDCl₃) δ, ppm: 10.64 (s, 1H), 8.58(s, 1H), 8.29-8.16 (m, 2H), 7.02-6.95 (m, 1H), 6.74-6.60 (m, 2H),5.88-5.82 (m, 1H), 4.66-4.62 (m, 1H), 4.38 (app t, J=11.0 Hz, 1H),4.38-4.28 (br m, 1H, overlapping with 4.38 ppm), 2.96-2.92 (m, 1H),2.26-2.23 (m, 1H); MS (EI) m/z 475.2 (MH)⁺.

(R)-2-(2-amino-5-fluorophenylamino)-6-chloro-9-(8-fluorochroman-4-yl)-7H-purin-8(9H)-one.To a round bottom flask containing(R)-6-chloro-2-(5-fluoro-2-nitrophenylamino)-9-(8-fluorochroman-4-yl)-7H-purin-8(9H)-one(310 mg, 0.65 mmol) was added glacial acetic acid (15 mL), water (37.5mL) and ethanol (75 mL), followed by iron powder (370 mg, 6.5 mmol, 10equiv.) and the resulting mixture heated at 90° C. for 15 min (HPLCmonitoring). The reaction mixture was left to cool down to roomtemperature, concentrated ammonium hydroxide solution (32 mL) was addedto bring the pH to basic, and the mixture was stirred for 10 min. Theaqueous layer was extracted with ethyl acetate (5×100 mL), the combinedorganic layers were washed with brine, dried (anhydrous Na₂SO₄), and thesolvent removed in vacuo to give the desired product as a pink solid(yield quantitative). This material was used in the next step withoutfurther purification. MS (EI) m/z 445.2 (MH)⁺.

6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-7H-purin-8(9H)-one.To a round bottom flask containing(R)-2-(2-amino-5-fluorophenylamino)-6-chloro-9-(8-fluorochroman-4-yl)-7H-purin-8(9H)-one(0.65 mmol) was added anhydrous methanol (45 mL), followed by anhydroustrimethylorthoformate (5 mL) and methane sulfonic acid (0.1 mL) and thereaction mixture was stirred under Ar at room temperature until it wascomplete (HPLC monitoring). Preparative HPLC purification, afterevaporation of solvent and drying, provided the desired product (184 mg,62% yield on two steps), as a white solid. ¹H NMR (300 MHz, CDCl₃) δ,ppm: 9.38 (br s, 1H), 9.01 (s, 1H), 7.84-7.79 (m, 1H), 7.64-7.60 (m,1H), 7.20-7.13 (m, 1H), 7.10-7.03 (m, 1H), 6.79-6.67 (m, 2H), 5.97-5.92(m, 1H), 4.68-4.62 (m, 1H), 4.51-4.40 (m, 1H), 2.95-2.83 (m, 1H),2.41-2.36 (m, 1H); MS (EI) m/z 454.9 (MH)⁺.

Method 1

General procedure for cross-coupling reactions of 6-chloropurinones witharylboronic acids. Ethanol (1 mL) was added to an argon-purged vialcontaining6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-7H-purin-8(9H)-one(9.2 mg, 0.02 mmol), aryl boronic acid (0.04 mmol, 2 equiv.), Pd(PPh₃)₄(2.3 mg, 0.002 mmol, 0.1 equiv.) and a 2M aqueous solution of Na₂CO₃(200 μL), and the mixture was heated for 30 min in the microwave oven at150° C. After cooling to ambient temperature, the reaction mixture wasdiluted with a 1/1 mixture of methanol/methylene chloride, filteredthrough a Nylon 0.45 μm filter and the filtrate concentrated in vacuo.Preparative HPLC purification of the residue afforded, after evaporationand drying, the desired compound.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-6-phenyl-7H-purin-8(9H)-one.White solid (4.8 mg, 48% yield). Prepared from phenyl boronic acid (5mg, 0.04 mmol, 2 equiv). ¹H NMR (300 MHz, CDCl₃) δ, ppm: 10.28 (br s,1H), 9.12 (s, 1H), 8.14-8.12 (m, 2H), 8.05-7.85 (m, 2H), 7.74-7.72 (m,3H), 7.28-7.26 (m, 1H), 7.18-7.16 (m, 1H), 6.88-6.84 (m, 2H), 6.14-6.12(m, 1H), 4.83-4.80 (m, 1H), 4.70-4.50 (m, 1H), 3.21-3.03 (m, 1H),2.60-2.42 (m, 1H); MS (EI) m/z 497.1 (MH)⁺.

Method 2

General procedure for displacement reactions of 6-chloropurinones withaliphatic amines. To 10 mg (0.022 mmol) of6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-7H-purin-8(9H)-onewas added 0.6 mL of 1-butanol, followed by an aliphatic amine (0.1 mL),and the reaction mixture was stirred at 110° C. for 1-2 h (HPLCmonitoring). Upon completion of the reaction, the mixture was left tocool to room temperature, was diluted with acetonitrile and filteredthrough a Nylon 0.45 μm filter. Preparative HPLC purification afforded,after solvent evaporation and drying, the desired product.

6-(cyclopropylmethylamino)-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-7H-purin-8(9H)-one.White solid (2.4 mg, 22% yield). Prepared using aminomethyl cyclopropane(0.1 mL). Reaction time 90 min. ¹H NMR (300 MHz, CD₃OD) δ, ppm: 9.01 (brs, 1H), 7.74-7.68 (m, 2H), 7.16-7.13 (m, 1H), 7.02-6.95 (m, 1H),6.77-6.69 (m, 2H), 5.88-5.83 (m, 1H), 4.79-4.61 (m, 1H), 4.45-4.38 (m,1H), 3.50 (d, J=6.9 Hz, 2H), 2.93-2.86 (m, 1H), 2.34-2.30 (m, 1H),1.29-1.21 (m, 1H), 0.64-0.61 (m, 2H), 0.39-0.36 (m, 2H); MS (EI) m/z490.1 (MH)⁺.

6-(azetidin-1-yl)-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-7H-purin-8(9H)-one.White solid, 70% yield. Prepared using azetidine. ¹H NMR (300 MHz,CDCl₃+CD₃OD) δ, ppm: 8.85 (br s, 1H), 7.75-7.67 (m, 2H), 7.13-6.90 (m,2H), 6.78-6.67 (m, 2H), 5.90-5.84 (m, 1H), 4.73-4.67 (m, 1H), 4.45-4.41(m, 5H), 3.00-2.95 (m, 1H), 2.63-2.55 (m, 2H), 2.36-2.30 (m, 1H); MS(EI) m/z 476.3 (MH)⁺.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-6-(3-hydroxyazetidin-1-yl)-7H-purin-8(9H)-one.White solid, 56% yield. Prepared using 3-hydroxyazetidine. ¹H NMR (300MHz, CDCl₃+CD₃OD) δ, ppm: 8.92 (br s, 1H), 7.66-7.60 (m, 2H), 7.09-7.02(m, 1H), 6.98-6.92 (m, 1H), 6.71-6.60 (m, 2H), 5.83-5.76 (m, 1H),4.78-4.62 (m, 1H), 4.59-4.52 (m, 3H), 4.40-4.32 (m, 1H), 4.14-4.07 (m,2H), 2.93-2.82 (m, 1H), 2.29-2.22 (m, 1H); MS (EI) m/z 492.3 (MH)⁺.

Method 3

General procedure for cross-coupling reactions of 6-chloropurinones withorganozinc halides. THF (1.5 mL) was added to an argon-purged vialcontaining6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-7H-purin-8(9H)-one(20 mg, 0.044 mmol) and Pd(PPh₃)₄ (2.5 mg, 0.02 mmol, 0.05 equiv.). Themixture was stirred at room temperature for 10 min, and then a 0.5 Msolution of an organozinc reagent in THF (132 μL, 0.066 mmol, 1.5equiv.) was added dropwise at room temperature. The stirring at roomtemperature was continued for 15 min, followed by stirring at 64° C. for1-3 days. The reaction mixture was left to cool to room temperature, thesolvent was removed in vacuo, acetonitrile was added to the residue andthe mixture filtered through a Nylon 0.45 μm filter. Preparative HPLCpurification afforded, after evaporation and drying, the desiredcompound.

4-(2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-8-oxo-8,9-dihydro-7H-purin-6-yl)butanenitrile.White solid, 16% yield. Prepared using a 0.5 M solution of3-cyanopropylzinc bromide in THF. ¹H NMR (300 MHz, CD₃OD) δ, ppm: 9.01(br s, 1H), 7.89-7.79 (m, 2H), 7.27-7.13 (m, 2H), 6.91 (br m, 2H),6.08-6.02 (m, 1H), 4.79-4.74 (m, 1H), 4.61-4.54 (m, 1H), 3.20 (t, J=7.2Hz, 2H), 3.04-2.97 (m, 1H), 2.77 (t, J=6.93 Hz, 2H), 2.52-2.40 (m, 1H,overlapping with 2.39 ppm), 2.39 (t, J=7.1 Hz, 2H); MS (EI) m/z 488.1(MH)⁺.

6-benzyl-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-((R)-8-fluorochroman-4-yl)-7H-purin-8(9H)-one.White solid, 49% yield. Prepared using a 0.5 M solution of benzylzincbromide in THF. ¹H NMR (300 MHz, CD₃OD) δ, ppm: 9.01 (br s, 1H),7.67-7.64 (m, 2H), 7.44-7.41 (m, 2H), 7.37-7.32 (m, 2H), 7.28-7.23 (m,1H), 7.17-7.11 (m, 1H), 7.03-6.97 (m, 1H), 6.77-6.69 (m, 2H), 5.93-5.87(m, 1H), 4.63-4.57 (m, 1H), 4.45-4.38 (m, 1H), 4.24 (s, 2H), 2.91-2.80(m, 1H), 2.39-2.33 (m, 1H); MS (EI) m/z 511.1 (MH)⁺.

General procedure for the selective displacement of the 4-chlorosubstituent in 2,4-dichloro-5-nitro-6-phenylpyrimidine with an amine. To200 mg of 2,4-dichloro-5-nitro-6-phenylpyrimidine (SPECS) (0.74 mmol) in2 mL anhydrous THF at −78° C. under Ar was added dropwise a solution ofan amine (0.74 mmol, 1.0 equiv.) and N,N-diisopropylethyl amine (284 μM,210 mg, 1.63 mmol, 2.2 equiv.) in 2 mL of THF. The reaction mixture wasstirred at −78° C. for 2 h, and then it was allowed to warm up to roomtemperature and stirred at ambient temperature for another 2 h (TLCmonitoring). The solvent was removed in vacuo and the residue purifiedby column chromatography to give the desired product.

2-chloro-5-nitro-6-phenyl-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine.Pale yellow solid. 104 mg, 42% yield. Synthesized using4-aminotetrahydropyran. Purification on silica gel, elution with 5/1hexanes/ethyl acetate. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 7.41-7.32(m, 5H), 4.35-4.29 (m, 1H), 4.02-3.92 (m, 2H), 3.51 (br t, 2H),2.01-1.94 (m, 2H), 1.66-1.56 (m, 2H).

(R)-2-chloro-N-(6-fluorochroman-4-yl)-5-nitro-6-phenylpyrimidin-4-amine.52% yield. Synthesized using (R)-6-fluorochroman-4-amine. Purificationon silica gel, elution with 7/3 hexanes/ethyl acetate. ¹H NMR (300 MHz,CDCl₃) δ, ppm: 7.62-7.43 (m, 6H), 7.01-6.93 (m, 2H), 6.88-6.82 (m, 1H),5.61-5.53 (m, 1H), 4.37-4.28 (m, 1H), 4.26-4.17 (m, 1H), 2.45-2.33 (m,1H), 2.22-2.13 (m, 1H); MS (EI) m/z 400.9 (MH)⁺.

2-(1H-benzo[d]imidazol-1-yl)-5-nitro-6-phenyl-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine.To 100 mg (0.3 mmol) of2-chloro-5-nitro-6-phenyl-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-aminein 5 mL of THF was added potassium carbonate (213 mg, 1.54 mmol, 5.1equiv.), followed by benzimidazole (106 mg, 0.9 mmol, 3 equiv.) and thereaction mixture was stirred at 70° C. After 2.5 h, heating wasdiscontinued; the reaction mixture was diluted with 70 mL of ethylacetate, was washed with water and dried (anhydrous MgSO₄). The solventwas removed in vacuo and the residue purified by column chromatography(silica gel, elution with 4/1 hexanes/ethyl acetate) to give the desiredproduct (48 mg, 38% yield). LC-MS: t_(R)=7.35 min, MS (EI) m/z 417.2(MH)⁺.

2-(1H-benzo[d]imidazol-1-yl)-N—((R)-6-fluorochroman-4-yl)-5-nitro-6-phenylpyrimidin-4-amine.Synthesized from(R)-2-chloro-N-(6-fluorochroman-4-yl)-5-nitro-6-phenylpyrimidin-4-amine.¹H NMR (300 MHz, CDCl₃) δ, ppm: 9.05 (s, 1H), 8.48-8.42 (m, 1H), 7.98(d, 1H), 7.87-7.82 (m, 1H), 7.67-7.61 (m, 2H), 7.58-7.48 (m, 3H),7.42-7.35 (m, 2H), 7.05-6.93 (m, 2H), 6.92-6.85 (m, 1H), 5.68-5.59 (m,1H), 4.42-4.31 (m, 1H), 4.30-4.19 (m, 1H), 2.51-2.41 (m, 1H), 2.40-2.29(m, 1H).

2-(1H-benzo[d]imidazol-1-yl)-6-phenyl-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-4,5-diamine.To 48 mg (0.11 mmol) of2-(1H-benzo[d]imidazol-1-yl)-5-nitro-6-phenyl-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-aminein 10 mL THF was added a solution of 300 mg Na₂S₂O₄ and 100 mg NaHCO₃ in30 mL of water, followed by 2 mL of methanol. The reaction mixture wasstirred at room temperature (HPLC monitoring). Upon completion of thereaction, the mixture was diluted with 70 mL of ethyl acetate, thelayers were separated, and the aqueous layer extracted with ethylacetate. The combined organic layers were washed with brine, dried(anhydrous MgSO₄), and the solvent removed in vacuo to give 30 mg (68%yield) of desired product, which was used in the next step withoutfurther purification. MS (EI) m/z 387.4 (MH)⁺.

2-(1H-benzo[d]imidazol-1-yl)-N⁴—((R)-6-fluorochroman-4-yl)-6-phenylpyrimidine-4,5-diamine.Synthesized from2-(1H-benzo[d]imidazol-1-yl)-N—((R)-6-fluorochroman-4-yl)-5-nitro-6-phenylpyrimidin-4-amine.MS (EI) m/z 453.3 (MH)⁺.

2-(1H-benzo[d]imidazol-1-yl)-6-phenyl-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.To 30 mg (0.078 mmol) of crude2-(1H-benzo[d]imidazol-1-yl)-6-phenyl-N⁴-(tetrahydro-2H-pyran-4-yl)pyrimidine-4,5-diaminein 3 mL THF was added CDI (38 mg, 0.233 mmol, 3 equiv.) and the reactionmixture was stirred at 50° C. for 18 h. The solvent was removed in vacuoand the residue purified by preparative TLC (silica gel, 5% methanol inmethylene chloride) to afford the desired product (9.5 mg, 30% yield).¹H NMR (300 MHz, CDCl₃) δ, ppm: 9.16 (br s, 1H), 8.68 (d, 1H), 8.06 (brm, 2H), 7.90 (d, 1H), 7.71-7.62 (m, 3H), 7.52-7.33 (m, 3H), 4.82-4.68(m, 1H), 4.31-4.20 (m, 2H), 3.64 (app t, 2H), 3.01-2.82 (m, 2H), 1.89(br d, 2H); MS (EI) m/z 413.2 (MH)⁺.

2-(1H-benzo[d]imidazol-1-yl)-9-((R)-6-fluorochroman-4-yl)-6-phenyl-7H-purin-8(9H)-one.26% yield. Synthesized from2-(1H-benzo[d]imidazol-1-yl)-N⁴—((R)-6-fluorochroman-4-yl)-6-phenylpyrimidine-4,5-diamine.¹H NMR (300 MHz, CDCl₃) δ, ppm: 10.72 (br s, 1H), 8.90 (s, 1H),8.18-8.04 (m, 3H), 7.82-7.77 (m, 1H), 7.63-7.54 (m, 3H), 7.36-7.28 (m,2H), 7.08-7.01 (m, 1H), 6.97-6.88 (m, 1H), 6.72-6.68 (m, 1H), 6.00-5.92(m, 1H), 4.63-4.56 (m, 1H), 4.36 (td, 1H), 3.05-2.90 (m, 1H), 2.40-2.26(m, 1H); MS (EI) m/z 479.1 (MH)⁺.

Ethyl2-(2-(tert-butoxycarbonyl)-5-fluorophenylamino)-5-nitro-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidine-4-carboxylate.To a solution of ethyl 2,6-dichloro-5-nitropyrimidine-4-carboxylate (2.0g, 7.5 mmol) in dry dioxane (30 mL) was added N,N-diisopropylethylamine(3.9 mL, 22.5 mmol) dropwise, at 0° C. To this solution was addedtetrahydro-2H-pyran-4-amine hydrochloride (1.04 g, 7.5 mmol) inaliquots, and the reaction mixture was slowly warmed to room temperatureand stirred for 30 min. It was cooled to 0° C. andN,N-diisopropylethylamine (3.9 mL, 22.5 mmol) was added dropwise,followed by tert-butyl-2-amino-4-fluorophenylcarbamate (1.6 g, 7.3 mmol)and the resulting mixture was heated to 60° C. for 6 h. The reactionmixture was cooled to room temperature and concentrated in vacuo. Theresidue was purified using column chromatography (silica gel, gradientelution with ethyl acetate in petroleum ether) to give the desiredproduct as a yellow solid. Yield: 530 mg, 14%. ¹H NMR (400 MHz, DMSO-d₆)δ, ppm: 9.91 (s, 1H), 8.72 (s, 1H), 8.59 (d, J=7.3 Hz, 1H), 7.61 (d,J=7.7 Hz, 1H), 7.53 (t, J=7.4 Hz, 1H), 7.05 (m, 1H), 4.36 (q, J=7.1 Hz,2H), 4.10 (m, 1H), 3.87 (d, J=11.0 Hz, 2H), 3.28 (t, J=12.2 Hz, 2H),1.81-1.67 (m, 4H), 1.45 (s, 9H), 1.30 (t, J=7.0 Hz, 3H); LCMS (EI) m/z521 (MH)⁺.

Ethyl2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-5-nitro-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidine-4-carboxylate.Ethyl2-(2-(tert-butoxycarbonyl)-5-fluorophenylamino)-5-nitro-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidine-4-carboxylate(530 mg, 1.02 mmol) was treated with a 30% (v) solution of TFA inmethylene chloride with stirring at room temperature. TLC monitoringshowed the reaction to be complete in 3 h. The solvent was evaporatedand the residue treated with a solution of trimethylorthoformate inmethanol (1:1 (v), 20 mL). After stirring at room temperature for 12 h,an orange solid formed. The orange solid was filtered and dried to givethe desired product (310 mg, 72% yield). ¹H NMR (400 MHz, DMSO-d₆) δ,ppm: 9.11 (s, 1H), 8.85 (d, J=7.6 Hz, 1H), 8.08 (d, J=2.5 Hz, 1H), 7.84(m, 1H), 7.33-7.27 (m, 1H), 4.75 (m, 1H), 4.60 (q, J=7.0 Hz, 2H), 3.96(d, J=11.0 Hz, 2H), 3.54 (t, J=7.3 Hz, 2H), 1.90-1.80 (m, 4H), 1.34 (t,J=7.1 Hz, 3H); LCMS (EI) m/z 431 (MH)⁺.

Ethyl5-amino-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidine-4-carboxylate.Hydrogenation of ethyl2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-5-nitro-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidine-4-carboxylate(310 mg, 0.717 mmol) was performed in ethanol, using Pd/C (10%), under apositive pressure of hydrogen. After 10 h, the catalyst was filtered,washed with portions of methylene chloride, and the combined filtrateconcentrated to give the desired amine as a white solid (220 mg, 78%yield). ¹H NMR (400 MHz, DMSO-d₆) δ, ppm: 8.97 (s, 1H), 8.47 (dd, J=2.6,2.2 Hz, 1H), 7.74 (m, 1H), 7.60 (d, J=8.9 Hz, 2H), 7.16 (m, 1H), 6.74(d, J=8.3 Hz, 1H), 4.37-4.32 (m, 3H), 3.94 (d, J=10.4 Hz, 2H), 3.55 (t,J=10.4 Hz, 2H), 2.01 (m, 2H), 1.58 (m, 2H), 1.42 (t, J=7.1 Hz, 3H); LCMS(EI) m/z 401 (MH)⁺.

Ethyl2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purine-6-carboxylate.To a solution of ethyl5-amino-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-6-(tetrahydro-2H-pyran-4-ylamino)pyrimidine-4-carboxylate(800 mg, 2.0 mmol) in dry THF (10 mL) was added carbonyl diimidazole(2.0 g) and the mixture was stirred at 60° C. for 48 h in a sealed tube.The solvent was evaporated and water (25 mL) was added to the residue.Filtration and drying of the white solid gave 600 mg (51% yield) ofdesired product. ¹H NMR (400 MHz, DMSO-d₆) δ, ppm: 11.89 (s, 1H), 9.09(s, 1H), 8.84 (dd, J=2.8, 2.4 Hz, 1H), 7.78 (dd, J=5.2, 5.2 Hz, 1H),7.23 (m, 1H), 4.63 (m, 1H), 4.49 (q, J=6.8 Hz, 2H), 4.40 (dd, J=4.0, 3.0Hz, 2H), 3.51 (t, J=11.6 Hz, 2H), 2.52 (m, 2H), 1.80 (d, J=2 Hz, 2H),1.41 (t, J=7.1 Hz, 3H); LCMS (EI) m/z 427 (MH)⁺.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purine-6-carboxylicacid. A solution of ethyl2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purine-6-carboxylate(600 mg, 1.41 mmol) in concentrated aqueous HCl (15 mL) was heated at100° C. for 6 h. The mixture was concentrated and the residue trituratedwith diethyl ether. The solid formed was filtered and dried to give 7 asan off white solid. Yield: 410 mg, 73%. ¹H NMR (400 MHz, DMSO-d₆) δ,ppm: 11.84 (s, 1H), 9.20 (s, 1H), 8.47 (dd, J=2.4, 2.4 Hz, 1H), 7.82(dd, J=5.2, 4.8 Hz, 1H), 7.26 (m, 1H), 4.63 (m, 1H), 4.04 (q, J=3.6 Hz,2H), 3.51 (t, J=11.6 Hz, 2H), 2.60-2.54 (m, 2H), 1.80 (d, J=9.8 Hz, 2H);LCMS (EI) m/z 399 (MH)⁺.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-6-(piperidine-1-carbonyl)-9-(tetrahydro-2H-pyran-4-yl)-7H-purin-8(9H)-one.To a solution of2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purine-6-carboxylicacid (25 mg, 0.063 mmol) in THF (2 mL) at 0° C. was added piperidine (8mg, 0.09 mmol) and triethyl amine (0.052 mL, 0.37 mmol), followed by asolution of triphenylphosphine (0.056 mL, 0.18 mmol) in ethyl acetate.The reaction mixture was allowed to reach room temperature and stirredfor 24 h. The solvent was evaporated and a saturated solution of sodiumbicarbonate was added to the residue. The solid formed was filtered anddried to give the desired compound as a white solid. Yield: 10 mg, 34%.¹H NMR (400 MHz, DMSO-d₆) δ, ppm: 11.89 (s, 1H), 8.20 (d, J=8.5 Hz, 1H),7.82 (dd, J=4.8, 4.8 Hz, 1H), 7.24 (t, J=6.94 Hz, 1H), 4.58 (m, 1H),4.03 (t, J=2.2 Hz, 2H), 3.68 (m, 2H), 3.68-3.48 (m, 4H), 2.64-2.55 (m,2H), 1.79 (d, J=11.6 Hz, 2H), 1.66 (m, 4H), 1.55 (m, 2H); LCMS (EI) m/z466 (MH)⁺.

6-chloropyrimidine-2,4,5-triamine hydrochloride was prepared asdescribed in WO 94/07892. 2,5-Diamino-4,6-dichloropyrimidine (10.8 g,60.3 mmol) and concentrated aqueous ammonia (30 mL) were heated togetherin a sealed tube at 105° C. for 18 h, then cooled to room temperature.2.34 g NaOH in water (10 mL) was added and the excess ammonia strippedout by concentrating to a low volume. Dilute HCl 2N (35 mL) and water(10 mL) were added and heated to dissolve, cooled to crystallize, thenfiltered to give the desired 6-chloropyrimidine-2,4,5-triaminehydrochloride (87% yield). MS (EI) m/z 160.1 (MH)⁺.

2-amino-6-chloro-7H-purin-8(9H)-one. To a solution of2,4,5-triamine-6-chloro-pyrimidine (6 g, 37 mmol) in 400 mL of anhydrousTHF was added 1,1-carbonyldiimidazole (37 g, 226 mmol) as a solid, inportions. The reaction mixture was stirred at room temperature for 2 h(completion of the reaction checked by HPLC and MS). The solvent wasremoved in vacuo. Water (300 mL) was added to the residue and themixture stirred at room temperature for 10 min. The solid formed wasfiltered under vacuum, and thoroughly dried to give 4.27 g (61% yield)of the desired product as a bright red solid. MS (EI) m/z 186.1 (MH)⁺.

3-(6-chloro-8-oxo-8,9-dihydro-7H-purin-2-ylamino)-4-nitrobenzonitrile.To an oven-dried microwave flask under Ar was added2-amino-6-chloro-7H-purin-8(9H)-one (0.64 g, 3.46 mmol, 1.0 equiv.)followed by anhydrous dioxane (10 mL) and anhydrous ethanol (4 mL);Cs₂CO₃ (Aldrich, 2.25 g, 6.92 mmol, 2 equiv.),3-bromo-4-nitro-benzonitrile (1.02 g, 4.5 mmol, 1.3 equiv.), Xantphos(Strem, 0.6 g, 1.04 mmol, 0.3 equiv.) and Pd₂(dba)₃ (Strem, 0.32 g, 0.35mmol, 0.1 equiv.) were all added as solids, and the mixture was stirredunder Ar, at room temperature for 3 min. The flask was capped and heatedin the microwave at 140° C. for 150 min (LC-MS shows reaction to becomplete). The reaction mixture was cooled to room temperature, thenconcentrated in vacuo to give a dark brown residue, which was purifiedby flash chromatography (silica gel, gradual elution 3/1 ethylacetate/hexanes, then 4/1 ethyl acetate/hexanes with 1% MeOH to 20%MeOH) to give the desired product as an orange solid in 46% yield. ¹HNMR (300 MHz, CD₃COOD) δ, ppm: 9.31 (d, 1H), 8.36 (d, 1H), 7.43 (dd,1H); MS (EI) m/z 332.0 (MH)⁺.

4-amino-3-(6-chloro-8-oxo-8,9-dihydro-7H-purin-2-ylamino)benzonitrile.To 3-(6-chloro-8-oxo-8,9-dihydro-7H-purin-2-ylamino)-4-nitrobenzonitrile(0.41 g, 1.23 mmol) was added glacial acetic acid (30 mL), water (75 mL)and ethanol (150 mL), followed by iron powder (Aldrich, 0.69 g, 12.3mmol, 10 equiv.) and the reaction mixture was stirred at 90° C. for 15min (LC-MS monitoring), when it became a clear dark orange solution. Itwas cooled to r.t., then concentrated NH₄OH (50 mL) was added to bringthe pH to 11. The reaction mixture was stirred at r.t. for 30 min.Repeated extractions into ethyl acetate, then into 25% isopropanol inCH₂Cl₂ (total volume 2 L), drying of the combined organic extracts, andin vacuo evaporation of the solvent provided the desired product as abrown solid, which was used in the next step without furtherpurification. MS (EI) m/z 302.1 (MH)⁺.

3-(6-chloro-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.To a solution of crude4-amino-3-(6-chloro-8-oxo-8,9-dihydro-7H-purin-2-ylamino)benzonitrile(1.23 mmol) in anhydrous DMSO (10 mL) was added methanol (150 mL),followed by anhydrous trimethylorthoformate (5 mL) and p-toluenesulfonic acid (100 mg) and the reaction mixture was stirred under Ar atroom temperature for 3 h (LC-MS monitoring). The solvent was removed invacuo and the residue redissolved in 7 mL of DMSO, then 40 mL of waterwere added. The pink solid formed was filtered under vacuo using a fineporosity Buchner filter funnel, washed with water, then dried in thevacuum oven at 60° C. to give 0.26 g of the desired product (67% yieldon 2 steps). ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.24 (s, 1H), 9.00(s, 1H), 7.91 (d, 1H), 7.69 (d, 1H); MS (EI) m/z 312.2 (MH)⁺.

General procedure for diversifying at the N9 position of the purinonesbased on a Mitsunobu reaction. A mixture of3-(6-substituted-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile(0.24 mmol, 1.0 equiv.) in CH₂Cl₂ (4 mL) and DMSO (1 mL), thecorresponding benzyl alcohol or primary aliphatic alcohol (0.42 mmol,1.75 equiv.) and resin-bound PS-triphenylphosphine (3 mmol/g, 172 mg,0.52 mmol, 2.2 equiv.) was treated with di-t-butylazodicarboxylate (0.42mmol, 1.75 equiv.) at room temperature. The mixture was allowed to stirat room temperature for 1-3 days. The reaction mixture was thenfiltered, the resin washed with CH₂Cl₂ (2×10 mL), methanol (10 mL) andCH₂Cl₂ (10 mL), and the combined filtrate and washings concentrated andsubjected to preparative HPLC purification to give the desiredN9-substituted purinone.

3-(6-chloro-8-oxo-9-(1-(pyridin-3-yl)ethyl)-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.Yellow oil (26% yield based on 73% conversion). Synthesized using1-pyridin-3-yl-ethanol and3-(6-chloro-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.¹H NMR (300 MHz, CD₃OD) δ, ppm: 9.27 (s, 1H), 9.20-9.00 (br m, 1H), 8.86(d, 1H), 8.88-8.77 (m, 1H, overlapping with 8.86 ppm), 8.68 (s, 1H),8.13 (br s, 1H), 7.87 (d, 1H), 7.68 (d, 1H), 6.17 (q, J=7.1 Hz, 1H),2.19 (d, J=7.3 Hz, 3H); MS (EI) m/z 417.1 (MH)⁺.

3-(6-(3,3-difluoroazetidin-1-yl)-8-oxo-9-(1-(pyridin-3-yl)ethyl)-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.White semi-solid. 57% yield. Synthesized from3-(6-chloro-8-oxo-9-(1-(pyridin-3-yl)ethyl)-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile(19 mg, 0.046 mmol) and 3,3-difluoroazetidine hydrochloride (3 equiv.)following the general procedure for displacement reactions of6-chloropurinones with aliphatic amines. ¹H NMR (300 MHz, CD₃OD) δ, ppm:9.27 (s, 1H), 9.20-9.00 (br m, 1H), 8.78 (d, 1H), 8.88-8.77 (m, 1H,overlapping with 8.78 ppm), 8.63 (s, 1H), 8.09 (br s, 1H), 7.88 (d, 1H),7.67 (d, 1H), 6.13 (q, J=7.1 Hz, 1H), 4.78 (t, 4H), 2.16 (d, J=7.1 Hz,3H); MS (EI) m/z 474.0 (MH)⁺.

3-(6-(3,3-difluoroazetidin-1-yl)-8-oxo-9-((tetrahydrofuran-3-yl)methyl)-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.White solid. 18% yield. Synthesized from3-(6-(3,3-difluoroazetidin-1-yl)-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile(0.12 mmol) and tetrahydro-3-furan-methanol following the generalprocedure for diversifying at the N9 position of the purinones based ona Mitsunobu reaction. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.22 (s,1H), 8.99 (s, 1H), 7.89 (d, 1H), 7.67 (d, 1H), 4.76 (t, 4H), 4.10-3.97(m, 3H), 3.90-3.77 (m, 3H), 3.06-2.93 (m, 1H), 2.22-2.07 (m, 1H),1.90-1.83 (m, 1H); IR (CHCl₃), cm⁻¹: 2979 (w), 1703 (s), 1367 (m), 1167(m); MS (EI) m/z 453.1 (MH)⁺.

6-chloro-N⁴-(trans-4-methoxycyclohexyl)pyrimidine-2,4,5-triamine.2,5-diamino-4,6-dichloropyrimidine (6 g, 33.5 mmol), trans-4-methoxycyclohexyl amine (4.32 g, 33.5 mmol, 1 equiv.), sodium bicarbonate (9.85g, 117.2 mmol, 3.5 equiv.) and 1-butanol (350 mL) were heated togetherat 150° C. in a sealed tube. After 3 days, when the reaction appeared tobe complete (by HPLC), the reaction mixture was cooled to roomtemperature and the solvent was removed in vacuo. The residue waspurified by flash chromatography (silica gel, gradual elution with 96/4methylene chloride/methanol to 90/10 methylene chloride/methanol) togive the desired product as a brown solid (69% yield). MS (EI) m/z 272(MH)⁺.

2-amino-6-chloro-9-(trans-4-methoxycyclohexyl)-7H-purin-8(9H)-one. To asolution of6-chloro-N⁴-(trans-4-methoxycyclohexyl)pyrimidine-2,4,5-triamine (5.5 g,20.36 mmol) in 300 mL of anhydrous THF (use oven dried glassware) wasadded 1,1-carbonyldiimidazole (newly opened bottle, 19.8 g, 122.2 mmol,6 equiv.) as a solid, in portions. The reaction mixture was stirred atroom temperature for 2 h (completion of the reaction checked by HPLC andMS). The solvent was removed in vacuo. Water (250 mL) was added to theresidue and the mixture stirred at room temperature for 10 min. Thesolid formed was filtered under vacuum, washed with cold water andthoroughly dried to give the desired product as a brown solid (67%yield), which was used in the next step without further purification. MS(EI) m/z 298 (MH)⁺.

3-(6-chloro-9-(trans-4-methoxycyclohexyl)-8-oxo-8,9-dihydro-7H-purin-2-ylamino)-4-nitrobenzonitrile.To an oven-dried 250 mL round bottom flask was added2-amino-6-chloro-9-(trans-4-methoxycyclohexyl)-7H-purin-8(9H)-one(crude, 3.65 g, 12.28 mmol, 1 equiv.) in anhydrous toluene (50 mL), thenfreshly grounded cesium carbonate (6 g, 18.43 mmol, 1.5 equiv.) withstirring at room temperature under Ar. Pd(OAc)₂ (1.24 g, 1.84 mmol, 0.15equiv.), racemic BINAP (2.67 g, 4.3 mmol, 0.35 equiv.) and3-bromo-4-nitro-benzonitrile (3.6 g, 15.96 mmol, 1.3 eq) were all addedas solids. The reaction was performed under a flow of Argon, withheating at 100° C. for two days. The reaction mixture was cooled to roomtemperature, then concentrated in vacuo and the resulting residue waspurified using flash chromatography (silica gel, gradient elution 10%EtOAc in hexanes to 50% EtOAc in hexanes) to give the desired product asan orange solid (50% yield). MS (EI) m/z 444 (MH)⁺.

6-chloro-2-(5-chloro-2-nitrophenylamino)-9-((trans)-4-methoxycyclohexyl)-7H-purin-8(9)-one.Synthesized using the above procedure. Yield 35%. MS (EI) m/z 453 (MH)⁺.

4-amino-3-(6-chloro-9-trans-4-methoxycyclohexyl)-8-oxo-8,9-dihydro-7H-purin-2-ylamino)benzonitrile.3-(6-chloro-9-(trans-4-methoxycyclohexyl)-8-oxo-8,9-dihydro-7H-purin-2-ylamino)-4-nitrobenzonitrile(1.3 g, 2.93 mmol) was taken in a 500 mL round bottom flask. To it wasadded iron powder (1.64 g, 29.3 mmol, 10 equiv.), followed byCH₃COOH:H₂O:EtOH (1:2.5:5 volume ratio) total volume 225 mL. Thereaction mixture was then heated to 90° C. with continuous stirring for30 minutes. Completion of the reaction was monitored by analytical HPLCand MS analysis (M⁺ 414). The reaction mixture was cooled to roomtemperature. Saturated NH₄OH was added to the cooled solution slowly,with stirring, until pH was 11-12. It was then diluted with EtOAc (250mL). The organic layer was separated and the aqueous layer extractedwith multiple portions of 200 mL of EtOAc until the aqueous layer wasfree of any desired compound (by HPLC). The organic layers werecombined, dried over MgSO₄ and concentrated to give a light brown solid(82% yield), which was used without purification in the next step. MS(EI) m/z 414 (MH)⁺.

2-(2-amino-5-chlorophenylamino)-6-chloro-9-((trans)-4-methoxycyclohexyl)-7H-purin-8(9H)-one.Synthesized using the above procedure. MS (EI) m/z 423 (MH)⁺.

3-(6-chloro-9-(trans-4-methoxycyclohexyl)-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.To a solution of4-amino-3-(6-chloro-9-trans-4-methoxycyclohexyl)-8-oxo-8,9-dihydro-7H-purin-2-ylamino)benzonitrile(1.0 g, 2.41 mmol) in 25 mL of anhydrous methanol was addedtrimethylorthoformate (1 mL, 10.89 mmol, 4.5 equiv.), followed by 8 to10 drops of methane sulfonic acid. The reaction mixture was stirred atroom temperature for 2 h. Completion of the reaction was checked by HPLCand MS. The reaction mixture was concentrated under high vacuum to givea dark brown solid. (97% crude yield). Preparative HPLC purification ofa 50 mg sample gave pure desired compound in 20% yield. ¹HNMR (300 MHz,CDCl₃+CD₃OD) δ, ppm: 9.22 (s, 1H), 8.96 (s, 1H), 7.98 (d, 1H), 7.72 (dd,1H), 4.54-4.40 (m, 1H), 3.58-3.40 (m, 1H, overlapping with 3.51 ppm),3.51 (s, 3H), 2.62-2.44 (m, 2H), 2.37 (br d, 2H), 2.00 (br d, 2H),1.59-1.41 (m, 2H); MS (EI) m/z 424.0 (MH)⁺.

6-chloro-2-(6-chloro-1Hbenzo[d]imidazol-1-yl)-9-((trans-4-methoxycyclohexyl)-7H-purin-8(9H)-one.Synthesized using the above procedure. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ,ppm: 9.03 (s, 1H), 8.58 (d, 1H), 7.72 (d, 1H), 7.38 (dd, 1H), 4.61-4.38(m, 1H), 3.58-3.48 (m, 1H), 3.45 (s, 3H), 2.66-2.45 (m, 2H), 2.33 (br d,2H), 1.99 (br d, 2H), 1.52-1.38 (m, 2H); MS (EI) m/z 433.1 (MH)⁺.

General Procedure for N-7 alkylation/methylation. To a solution of3-(6-chloro-9-(trans-4-methoxycyclohexyl)-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile(1.65 mmol) in CH₃CN (25 mL) was added polystyrene supported BEMP(2-tert.butylimino-2-diethylamino-1,3-dimethyl-perhydro-1,3,2-diazaphosphorine(Fluka, loading 2.2 mmol/g) (4 equiv.), followed by alkyliodide/iodomethane (6 equiv.). The reaction mixture was stirred at roomtemperature for 1 h. Completion of the reaction mixture was checked byHPLC and MS. The reaction mixture was filtered and the resin was washedwith CH₃CN (10 mL×2) and MeOH (10 mL×2). The washings and the filtratewere combined and concentrated in vacuo to give the desired compound.

3-(6-chloro-9-(trans-4-methoxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.Dark brown solid (76% crude yield). Synthesized using3-(6-chloro-9-(trans-4-methoxycyclohexyl)-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile(0.7 g, 1.65 mmol) and iodomethane (0.61 mL, 10 mmol, 6 equiv.). MS (EI)m/z 438 (MH)⁺.

6-Chloro-2-(6-chloro-1H-benzo[d]imidazol-1-yl)-9-((trans)4-methoxycyclohexyl)-7-methyl-7H-purin-8(9H)-one.Synthesized using the above procedure. MS (EI) m/z 447 (MH)⁺.

Typical Procedure for Selective Demethylation

3-(6-chloro-9-(trans-4-hydroxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]/imidazole-5-carbonitrile.To an oven dried flask under Argon flow was added3-(6-chloro-9-(trans-4-methoxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile(0.55 g, 1.25 mmol) in chloroform, followed by iodotrimethylsilane (0.1mL, 1.25 mmol, 1 equiv.) The reaction mixture was stirred at roomtemperature for 4 h. Upon completion (HPLC monitoring), the reaction wasslowly quenched with water. The organic layer was separated and theaqueous layer extracted with CHCl₃ (15 ml×3) and with EtOAc (15 mL×3).The combined organic layers were dried (MgSO₄), and concentrated invacuo to give a dark solid (66% crude yield). MS (EI) m/z 424 (MH)⁺.

6-chloro-2-(6-chloro-1H-benzo[d]imidazol-1-yl)-9-((trans)-4-hydroxycyclohexyl)-7-methyl-7-purin-8(9H)-one.Synthesized using the above procedure. MS (EI) m/z 433 (MH)⁺.

Typical procedure for cross-coupling reactions of 6-chloropurinones with(hetero)arylboronic acids

3-(9-((trans-4-hydroxycyclohexyl)-7-methyl-8-oxo-6-(pyridin-4-yl)8,9-dihydro-7H-purin-2yl)3-H-benzo[d]imidazole-5-carbonitrile.Ethanol (3 mL) was added to an argon-purged vial containing3-(6-chloro-9-(trans-4-hydroxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile(100 mg, 0.236 mmol), pyridine-4-boronic acid pinacol ester (97 mg, 0.47mmol, 2 equiv.), Pd(PPh₃)₄ (27 mg, 0.0236 mmol, 0.1 equiv.) and a 2Maqueous solution of Na₂CO₃ (200 μL). The mixture was heated for 30 minin the microwave oven at 150° C. After cooling to ambient temperature,the reaction mixture was diluted with ethanol, filtered through a Nylon0.45 μm filter and the filtrate concentrated in vacuo. Preparative HPLCpurification of the residue afforded, after evaporation and drying, thedesired compound (TFA salt). ¹HNMR (300 MHz, CDCl₃) δ, ppm: 9.30 (s,1H), 9.04 (br s, 1H), 9.04-9.00 (br d, 2H, overlapping with 9.04 ppm),8.05 (d, 1H), 7.76 (dd, 1H), 7.68 (br d, 2H), 4.74-4.60 (m, 1H),4.20-4.08 (m, 1H), 3.33 (s, 3H), 2.84-2.67 (m, 2H), 2.42-2.31 (m, 2H),2.17-2.04 (m, 2H), 1.82-1.64 (m, 2H), 1.36 (br s, 1H); Structureconfirmed by nOe and FTIR; MS (EI) m/z 467.1 (MH)⁺.

Typical procedure for the Displacement of Chlorine in 6-Chloropurinoneswith Amines

2-(6-chloro-1H-benzo[d]imidazole-1-yl)-6-(3,3-difluoroazetidin-1-yl)-9-(trans-4-hydroxycyclohexyl)-7-methyl-7H-purin-8(9H)-one.6-Chloro-2-(6-chloro-1H-benzo[d]imidazole-1-yl-9-((trans-4-hydroxycyclohexyl)-7-methyl-7H-purin-8(9H)-one(166 mg, 0.38 mmol) was taken in a scintillation vial with 1-butanol (2mL), DMSO (1 mL) and DIEA (200 μL). To it was then added3,3-difluoroazetidine hydrochloride (220 mg, 1.71 mmol, 4.5 equiv.) Thereaction mixture was stirred at 110° C. for 18 h. The solvent was thenremoved under high vacuum. The residue was dissolved in MeOH andfiltered. Preparative HPLC purification of the residue afforded, afterevaporation and drying, the desired compound. White solid (no salt).¹HNMR (300 MHz, CDCl₃) δ, ppm: 8.92 (s, 1H), 8.53 (d, 1H), 7.76 (d, 1H),7.34 (dd, 1H), 4.71 (t, 4H), 4.53-4.41 (m, 1H), 4.07-3.93 (m, 1H), 3.55(s, 3H), 2.69-2.51 (m, 2H), 2.27-2.15 (m, 2H), 1.97-1.87 (m, 2H),1.64-1.51 (m, 2H, overlapping with 1.56 ppm), 1.56 (br s, 1H); Structureconfirmed by nOe, NOESY and FTIR; MS (EI) m/z 490.1 (MH)⁺.

3-(6-(3,3-difluoroazetidin-1-yl)-9-(trans-4-hydroxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.Synthesized using the procedure above. ¹HNMR (300 MHz, CDCl₃+CD₃OD) δ,ppm: 9.15 (s, 1H), 8.91 (s, 1H), 7.91 (d, 1H), 7.66 (dd, 1H), 4.74 (t,4H), 4.51-4.39 (m, 1H), 3.98-3.83 (m, 1H), 3.56 (s, 3H), 2.64-2.46 (m,2H), 2.24-2.13 (m, 2H), 1.97-1.87 (m, 2H), 1.62-1.46 (m, 2H); Structureconfirmed by nOe; MS (EI) m/z 481.2 (MH)⁺.

3-(6-(3,3-difluoroazetidin-1-yl)-9-(trans-4-hydroxycyclohexyl)-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrile.Synthesized from3-(6-chloro-9-(trans-4-methoxycyclohexyl)-8-oxo-8,9-dihydro-7H-purin-2-yl)-3H-benzo[d]imidazole-5-carbonitrileby demethylation, followed by displacement of chlorine in the6-chloropurinone with 3,3-difluoroazetidine. ¹HNMR (300 MHz,CDCl₃+CD₃OD) δ, ppm: 9.12 (s, 1H), 8.92 (s, 1H), 7.90 (d, 1H), 7.64 (dd,1H), 4.71 (t, 4H), 4.46-4.32 (m, 1H), 3.98-3.83 (m, 1H), 2.64-2.46 (m,2H), 2.24-2.15 (m, 2H), 1.97-1.87 (m, 2H), 1.62-1.46 (m, 2H); MS (EI)m/z 467.0 (MH)⁺.

Trans-4-(2,5-diamino-6-chloropyrimidin-4-ylamino)cyclohexanol.2,5-diamino-4,6-dichloropyrimidine (10.5 g, 58.6 mmol),trans-4-aminocyclohexanol (6.75 g, 58.6 mmol, 1 equiv.), sodiumbicarbonate (17.2 g, 205 mmol, 3.5 equiv.) and 1-butanol (210 mL) wereheated together at 150° C. in a sealed tube. After 3 days, when thereaction appeared to be complete (by LCMS), the reaction mixture wascooled to room temperature and the solvent was removed in vacuo. Water(150 mL) was added to the residue with stirring at room temperature for30 min, and the dark red solid filtered under vacuo to give, afterdrying, 10 g (66% yield) of the desired product. ¹H NMR (300 MHz,d₆-DMSO) δ, ppm: 6.14 (d, 1H), 5.57 (s, 2H), 4.54 (d, 1H), 3.90 (s, 2H),3.81-3.76 (m, 1H), 3.40-3.32 (m, 1H), 1.86-1.82 (m, 4H), 1.28-1.20 (m,4H); MS (EI) m/z 258.1 (MH)⁺.

Trans-4-(2-amino-6-chloro-8-oxo-7,8-dihydropurin-9-yl)cyclohexyl-1H-imidazole-1-carboxylate.To a solution oftrans-4-(2,5-diamino-6-chloropyrimidin-4-ylamino)cyclohexanol (8.7 g,33.9 mmol) in 430 mL of anhydrous THF (use oven dried glassware) wasadded 1,1-carbonyldiimidazole (newly opened bottle, 33 g, 203 mmol) as asolid, in portions. The reaction mixture was stirred at room temperaturefor 4 h (completion of the reaction checked by HPLC and MS). The solventwas removed in vacuo. Water (1 L) was added to the residue and themixture stirred at room temperature for 18 h. The solid was filteredunder vacuum. The procedure was repeated twice and the resulting solidwas thoroughly dried to give 10.6 g (83% yield) of the desired productas a pale brown solid. ¹H NMR (300 MHz, d₆-DMSO) δ, ppm: 11.3 (br s,1H), 8.27 (br s, 1H), 7.60 (br s, 1H), 7.08 (br s, 1H), 6.58 (s, 2H),4.92-4.84 (m, 1H), 4.22-4.13 (m, 1H), 2.45-2.37 (m, 2H), 2.23-2.20 (m,2H), 1.84-1.61 (m, 4H); MS (EI) m/z 378.4 (minor, MH)⁺, 266.1(fragment).

Trans-4-(2-amino-6-chloro-8-oxo-7,8-dihydropurin-9-yl)cyclohexyl methylcarbonate. To a suspension oftrans-4-(2-amino-6-chloro-8-oxo-7,8-dihydropurin-9-yl)cyclohexyl-1H-imidazole-1-carboxylate(10.6 g, 28.1 mmol) in anhydrous methanol (400 mL) was added potassiumcarbonate (1.1 g, 10% wt) with stirring at room temperature. After 15 h,the reaction was found to be complete by LC-MS. The solvent was removedin vacuo, and water (1 L) was added with stirring at room temperature.Filtration under vacuum gave the desired product as a pink solid (yieldquantitative). MS (EI) m/z 342.1 (MH)⁺.

2-amino-6-chloro-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one.Trans-4-(2-Amino-6-chloro-8-oxo-7,8-dihydropurin-9-yl)cyclohexyl methylcarbonate (1.43 g) was dissolved in 80 mL of an aqueous solution 0.5 NKOH and the reaction mixture heated at reflux for 30 min (LC-MS analysisindicated reaction to be complete). The reaction mixture was then cooledin an ice bath, and its pH was brought to 7 by drop wise addition of aconcentrated aqueous solution of HCl. A light pink solid precipitated.It was filtered under vacuum, thoroughly dried in vacuum oven at 60° C.,to give the desired product in 86% yield. ¹H NMR (300 MHz, CDCl₃+CD₃OD)δ, ppm: 8.48 (br s, 1H), 7.73 (s, 2H), 4.26-4.18 (m, 1H), 3.70-3.61 (m,1H), 2.53-2.40 (m, 2H), 2.10-2.06 (m, 2H), 1.81-1.76 (m, 2H), 1.48-1.37(m, 2H); MS (EI) m/z 284.1 (MH)⁺.

Alternative one step procedure for2-amino-6-chloro-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one.Trans-4-(2-amino-6-chloro-8-oxo-7,8-dihydropurin-9-yl)cyclohexyl-1H-imidazole-1-carboxylate(63.9 mmol) was suspended in an aqueous solution 0.5 N KOH (620 mL) withstirring at room temperature. The suspension gradually became a dark redsolution. After 30 min, the reaction was found to be complete by LC-MS.The pH of the reaction mixture was brought to 7 by dropwise addition ofa concentrated aqueous solution of HCl. A light pink solid precipitated.The neutralized reaction mixture was diluted with water up to a volumeof 2 L, with stirring at room temperature for 10 min. The solid wasfiltered under vacuum, thoroughly dried in vacuum oven at 60° C., togive the desired product (17.08 g, 94% yield). ¹H NMR (300 MHz,CDCl₃+CD₃OD) δ, ppm: 8.48 (br s, 1H), 7.73 (s, 2H), 4.26-4.18 (m, 1H),3.70-3.61 (m, 1H), 2.53-2.40 (m, 2H), 2.10-2.06 (m, 2H), 1.81-1.76 (m,2H), 1.48-1.37 (m, 2H); MS (EI) m/z 284.1 (MH)⁺.

6-chloro-2-(5-fluoro-2-nitrophenylamino)-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one.All glassware was dried in vacuum oven at 60° C. for 1 day prior toreaction. Finely ground cesium carbonate and finely ground2-amino-6-chloro-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one weredried at 60° C. under high vacuum for one day prior to experiment. To anoven-dried 2-neck flask under Ar was added2-amino-6-chloro-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one (6.78g, 23.9 mmol), cesium carbonate (11.71 g, 35.9 mmol, 1.5 equiv.) andanhydrous toluene (100 mL) and the mixture was stirred at roomtemperature for 15 min. Then, Pd(OAc)₂ (2.41 g, 3.59 mmol, 0.15 equiv.),racemic BINAP (5.21 g, 8.36 mmol, 0.35 equiv.) and1-bromo-5-fluoro-2-nitrobenzene (6.84 g, 31.07 mmol, 1.3 equiv.) wereadded as solids, under Ar, and the reaction mixture was stirred at roomtemperature for 15 min, then stirred at 100° C. for 2 days (LC-MS showsratio of desired product/starting material approx. 8/1). The reactionmixture was cooled to room temperature, and the solvent removed invacuo. The dark brown residue was dissolved in warm CH₃COOH/EtOH (60mL/60 mL), and then water (700 mL) was added. A dark yellow solidprecipitated and was filtered under vacuum. A small portion of the darkyellow solid was purified by flash chromatography (silica gel, gradientelution with ethyl acetate/hexanes 1/1 to 3/1, then up to 20% methanolin 3/1 ethyl acetate/hexanes) to provide the desired compound as anorange solid. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 10.67 (br s, 1H),8.80 (dd, 1H), 8.37 (dd, 1H), 6.85-6.80 (m, 1H), 4.39-4.30 (m, 1H),3.81-3.73 (m, 1H), 2.67-2.44 (m, 2H), 2.18-2.14 (m, 2H), 1.92-1.88 (m,2H), 1.56-1.41 (m, 2H); MS (EI) m/z 423.3 (MH)⁺.

2-(2-amino-5-fluorophenylamino)-6-chloro-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one.6-chloro-2-(5-fluoro-2-nitrophenylamino)-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one(13.5 mg, 0.03 mmol) was dissolved in CH₃COOH:H₂O:EtOH (1 mL:2.5 mL:5mL) and iron powder (18 mg, 0.32 mmol, 10 equiv.) was added. Thereaction mixture was then heated to 90° C. with continuous stirring for15 minutes. Completion of the reaction was monitored by analytical HPLCand MS analysis. The reaction mixture was cooled to room temperature.Saturated NH₄OH was added to the cooled solution slowly, with stirring,until pH was 11-12. It was then diluted with EtOAc (15 mL). The organiclayer was separated, and the aqueous layer extracted with EtOAc (3×15mL), the combined organic layers were washed with brine (1×15 mL), driedover MgSO₄ and concentrated to give a white solid (12 mg, 96% yield),which was used without purification in the next step. MS (EI) m/z 393(MH)⁺.

6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one.To a flask containing crude2-(2-amino-5-fluorophenylamino)-6-chloro-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one(12 mg, 0.03 mmol) was added anhydrous methanol (3 mL), followed byanhydrous trimethylorthoformate (0.5 mL) and p-toluenesulfonic acid(catalytic) and the reaction mixture was stirred under Ar at roomtemperature for 18 h (HPLC monitoring). A solution 10% acetonitrile inwater (total volume 10 mL) was added and a pale yellow solidprecipitated. Filtration under vacuum provided the desired product as apale yellow solid (10.6 mg, 88% yield). ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ,ppm: 9.07 (br s, 1H), 8.30 (br d, 1H), 7.82-7.70 (m, 1H), 7.27-7.15 (m,1H), 4.48-4.42 (m, 1H), 3.88-3.80 (m, 1H), 2.67-2.50 (m, 2H), 2.33-2.18(m, 2H), 2.10-1.95 (m, 2H), 1.70-1.56 (m, 2H); MS (EI) m/z 403.1 (MH)⁺.

6-(3,3-difluoroazetidin-1-yl)-2-(6-fluoro-3H-benzo[d]imidazol-1-yl)-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one.Pale yellow solid. TFA salt. Synthesized following the general procedurefor displacement reactions of 6-chloropurinones with aliphatic amines,using 3,3-difluoroazetidine. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.07(br s, 1H), 8.42-8.25 (br m, 1H), 7.82-7.70 (m, 1H), 7.15 (br d, 1H),4.71 (t, 4H), 4.48-4.38 (m, 1H), 3.88-3.80 (m, 1H), 2.67-2.50 (m, 2H),2.27-2.15 (m, 2H), 1.97-1.87 (m, 2H), 1.62-1.50 (m, 2H); MS (EI) m/z460.2 (MH)⁺.

Typical Procedure for Tetrahydropyranylation of Alcohols

6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(trans-4-(tetrahydro-2H-pyran-2-yloxy)cyclohexyl)-7H-purin-8(9H)-one.A solution of6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one(0.8 mmol) and dihydropyran (15 equiv.) in anhydrous chloroform/THF (30mL/15 mL) containing p-TsOH (0.1 equiv.) was stirred at 50° C. for 1 day(LCMS indicative of complete reaction). The reaction mixture wasconcentrated in vacuo and the residue purified by column chromatographyon neutral alumina to give the desired product. MS (EI) m/z 487.3 (MH)⁺.

Typical Procedure for N7-Methylation/Alkylation

6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-7-methyl-9-(trans-4-(tetrahydro-2H-pyran-2-yloxy)cyclohexyl)-7H-purin-8(9H)-one.To a solution of6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(trans-4-(tetrahydro-2H-pyran-2-yloxy)cyclohexyl)-7H-purin-8(9H)-one(0.06 mmol) in anhydrous CH₃CN (10 mL) were added BEMP on polystyrene(loading 2.2 mmol/g, 4 equiv.) and iodomethane (primary alkyliodide/benzyl iodide or bromide to be used for alkylation) (6 equiv.)with stirring at room temperature. After 1 h, LCMS showed reaction to becomplete. The reaction mixture was filtered under vacuum, the resinwashed with CH₃CN and methanol, and the combined filtrate and washingswere concentrated in vacuo to give the desired product as a pale yellowsolid, which was used in the next step without further purification. MS(EI) m/z 501.2 (MH)⁺.

Typical Procedure for the Hydrolysis of Tetrahydropyranyl Ethers

6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(trans-4-hydroxycyclohexyl)-7-methyl-7H-purin-8(9H)-one.A solution of6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-7-methyl-9-(trans-4-(tetrahydro-2H-pyran-2-yloxy)cyclohexyl)-7H-purin-8(9H)-one(0.06 mmol) and PPTS (5 mg) in ethanol/water (4.5 mL/0.5 mL) was stirredat reflux for 1 h. The solvent was evaporated in vacuo and the residuecould be purified by chromatography or used in the next step withoutpurification. MS (EI) m/z 417.0 (MH)⁺.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-6-((S)-3-fluoropyrrolidin-1-yl)-9-(trans-4-hydroxycyclohexyl)-7-methyl-7H-purin-8(9H)-one.Light brown solid. TFA salt. Synthesized from6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(trans-4-hydroxycyclohexyl)-7-methyl-7H-purin-8(9H)-oneand S-(+)-3-fluoropyrrolidine hydrochloride, following the generalprocedure for displacement reactions of 6-chloropurinones with aliphaticamines. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 8.99 (s, 1H), 8.27 (dd,1H), 7.74 (dd, 1H), 7.15 (td, 1H), 5.47 (d, J=52.4 Hz, 1H), 4.51-4.42(m, 1H), 4.19-3.83 (overlap of 5H), 3.65 (s, 3H), 2.68-2.42 (overlap of3H), 2.29-2.03 (overlap of 3H), 1.97-1.90 (m, 2H), 1.62-1.49 (m, 2H); MS(EI) m/z 470.3 (MH)⁺.

6-(azetidin-1-yl)-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(trans-4-hydroxycyclohexyl)-7-methyl-7H-purin-8(9H)-one.White solid. ¹H NMR (300 MHz, CDCl₃+CD₃OD) δ, ppm: 9.05 (s, 1H), 8.32(br d, 1H), 7.80-7.68 (br m, 1H), 7.20-7.08 (m, 1H), 4.51-4.42 (m, 1H),4.34 (t, 4H), 3.93-3.83 (m, 1H), 3.55 (s, 3H), 2.68-2.50 (overlap of4H), 2.25-2.11 (m, 2H), 1.97-1.83 (m, 2H), 1.62-1.49 (m, 2H); MS (EI)m/z 438.3 (MH)⁺.

2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-6-hydroxy-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one.An oven dried vial containing a stirbar was charged with Pd₂ dba₃ (2 mg,0.002 mmol, 0.02 equiv.),2-di-t-butylphosphino-2′,4′,6′-tri-isopropyl-1,1′-diphenyl (Strem, 3.5mg, 0.008 mmol, 0.08 equiv.), KOH (17 mg, 0.3 mmol, 3 equiv.) and6-chloro-2-(6-fluoro-1H-benzo[d]imidazol-1-yl)-9-(trans-4-hydroxycyclohexyl)-7H-purin-8(9H)-one(40 mg, 0.1 mmol, 1 equiv.), followed by 1,4-dioxane (0.3 mL) anddegassed water (0.3 mL). A stream of argon was passed through thereaction mixture for 3 min; the vial was then closed and heated at 100°C. After 16 h, the reaction was found to be complete. Preparative HPLCchromatography provided, after evaporation of solvent, the desiredproduct as a white solid. ¹H NMR (300 MHz, CD₃OD+CDCl₃) δ, ppm: 9.04 (s,1H), 8.45 (dd, 1H), 7.80-7.65 (m, 1H), 7.15-7.08 (m, 1H), 4.42-4.30 (m,1H), 3.84-3.77 (m, 1H), 2.70-2.60 (m, 2H), 2.25-2.11 (m, 2H), 1.97-1.83(m, 2H), 1.62-1.40 (m, 2H); MS (EI) m/z 385.3 (MH)⁺.

Jak3 Kinase Assay

Human Jak3 cDNA was amplified by PCR. A fragment encoding the catalyticdomain of Jak3 (508aa to 1124aa) was ligated with GST at 5′ end. Thisfused GST-Jak3 DNA fragment was cloned into the EcoRI site of the donorplasmid pFastBac 1 (Life Technologies #10359-016). The transformation,transposition, and transfection of insect cells (Sf9) were performedaccording to the manufacture's instructions. The cell lysate containingrecombinant GST-Jak3 was used in the kinase assay. Anti-GST antibody (10μg/ml, Sigma #G1417) was coated onto a 384-well plate at 4° C.overnight. Cell lysate containing GST-Jak3 (1:100 dilution) was added tothe anti-GST coated plates, and GST-Jak3 was captured by immobilizedanti-GST antibody. Testing compounds and substrate mix (50 mM HEPES, pH7, 0.5 mM Na₃VO₄, 25 mM MgCl₂, 1 mM DTT, 0.005% BSA, 1 μM ATP, and 4.5μg/ml biotinyl poly-Glu,Ala,Tyr) were added to the plate to initiate thereaction. After a 60-min incubation, the reaction was stopped by 4 mMEDTA, and phosphorylation of biotinyl poly-Glu,Ala,Tyr was detectedusing 17 μg/ml Cy5-streptavidin (Amersham, #PA92005) and 2.7 μg/mlEuropium-conjugated anti-phosphotyrosine antibody (PerkinElmer #AD0069)using homogeneous time-resolved fluorescence (HTRF) technology.

Jak3 Cellular Assay

The mouse F7 pre-B lymphocyte cell line was used for the cellular Jak3assay. Human IL-2Rβc cDNA is stably expressed in F7 cells (Kawahara etal., 1995). F7 cells were maintained in RPMI 1640 medium supplementedwith 10% fetal bovine serum plus IL-3. Cells (30,000 cells/well) inserum-free medium were seeded in 96-well plates for the cellproliferation assay. Testing compounds were added to cells, followed bythe addition of IL-2 (final 20 ng/ml). After a 24-h incubation, thenumber of viable cells was determined by the CellTiter-Glo LuminescentCell Viability Assay kit (Promega, #G7573) according to themanufacturer's instructions.

IL-2-Induced IFN-γ Production in the Mouse

Administration of IL-2 leads to an increase in serum IFN-γ in the mousedue to NK secretion of the cytokine (Thornton S, Kuhn K A, Finkelman F Dand Hirsch R. NK cells secrete high levels of IFN-γ in response to invivo administration of IL-2. Eur J Immunol 2001 31:3355-3360). Theexperiment is carried out essentially according to the protocol inThornton et al. and the test compounds are administered in order todetermine the level of inhibition attained. In summary, female BALB/cmice are fasted for 12-18 hours before a study but have free access towater at all times. Test compounds are administered by gavage one hourbefore intraperitoneal injection of IL-2 and capture antibody. Attermination of the studies, the mice are sacrificed by carbon dioxideinhalation, terminal blood samples are collected by cardiac puncture andserum is generated. Serum is stored frozen until assayed for IFN-γ, asdescribed by the kit manufacturer (BD Pharmingen™, San Diego, Calif.).

Some comparative examples are shown below. All of the IC₅₀'s for Jak3are below 10 μM.

C6 Substituted Purines and Purinones

Jak3 Kinase, IC50 Synthetic CHEMISTRY representation route

1 A2

1 I

1 A2

1 I

1 A2

1 A2

1 A2

1 J

1 A2

1 A2

1 A2

1 E2

1 I

1 A2

1 A2

1 J

1 A2

1 A2

1 J

1 I

1 A2

1 J

1 A2

1 A2

1 A2

1 J

1 I

1 A2

1 A2

1 A2

1 A2

1 I

1 A2

1 A2

1 A2

1 J

1 A2

1 A2

1 H

1 C

1 A2

1 A2

1 A2

1 A2

1 A2

1 A2

1 A1

1 A2

1 A1

1 D

1 E2

1 C

1 A2

1 A2

1 A2

1 A2

1 A2

1 A2

1 J

1 I

1 A2

1 A2

1 I

1 H

1 A2

1 G

1 E3

1 H

1 A2

1 A1

1 AB

1 A2

1 H

1 A2

1 G

1 G

1 G

1 J

1 H

1 A1

1 A2

1 I

1 A1

1 A2

1 A4

1 G

1 H

1 A2

1 E

1 G

1 A1

1 AB

1 A4

1 A2

1 J

1 G

1 A4

1 A1

1 B

1 A1

1 A1

1 G

1 A1

1 A1

2 C1

2 G

2 A1

2 G

2 G

2 A4

2 G

2 E3

2 G

2 A4

2 I

2 E1

2 A2

2 E1

2 A2

2 A1

2 A3

2 E2

2 E1

2 E3

2 A2

2 F

2 A1

2 E1

2 E2

2 E3

2 B

2 A1

2 E3

2 E3

2 B

3 A2In the foregoing table, an IC₅₀ less than 100 nM is represented as 1; anIC₅₀ between 100 nM and 1 μM is represented as 2; and an IC₅₀ greaterthan 1 μM is represented as 3.

1. A compound of formula I or II

wherein Q is selected from the group consisting of CX and nitrogen; X isselected from the group consisting of hydrogen, halogen, and anelectron-withdrawing group; A is chosen from the group consisting of H,(C₁-C₆) alkyl, heteroaryl, and aryl; R¹ is selected from the groupconsisting of halogen, CN, (C₂-C₆) alkyl, substituted (C₁-C₆) alkyl,aryl, substituted aryl, azetidinyl, piperidinyl, piperazinyl, pyridinyl,pyrimidinyl, morpholinyl, thiomorpholinyl, pyrrolyl, pyrazolyl, indolyl,isoquinolinyl, azepinyl, diazepinyl, cyclobutyl, substituted azetidinyl,substituted piperidinyl, substituted piperazinyl, substituted pyridinyl,substituted pyrimidinyl, substituted morpholinyl, substitutedthiomorpholinyl, substituted pyrrolyl, substituted pyrazolyl,substituted indolyl, substituted isoquinolinyl, substituted azepinyl,substituted diazepinyl, substituted cyclobutyl, and —V—R⁷; R² and R³ areselected independently for each occurrence of (CR²R³) from the groupconsisting of hydrogen and (C₁-C₆) alkyl; R⁴ is selected from a groupconsisting of alkyl, OH, alkoxy, pyranyl, benzopyranyl, furanyl,pyridinyl, cyclohexyl, aryl, substituted alkyl, substituted pyranyl,substituted benzopyranyl, substituted furanyl, substituted pyridinyl,substituted cyclohexyl, and substituted aryl; R⁷ is chosen from H,(C₁-C₆) alkyl, substituted (C₁-C₆) alkyl, aryl, substituted aryl,pyridinyl, piperidinyl, furanyl, oxadiazolyl, azetidinyl, cyclopropyl,cyclobutyl, cyclopentyl, and substituted pyridinyl, substitutedpiperidinyl, substituted furanyl, substituted oxadiazolyl, substitutedazetidinyl, substituted cyclopropyl, substituted cyclobutyl, substitutedcyclopentyl; V is chosen from —C(═O)O—, —C(═O)NR⁸—, —O—, and —NR⁸—; R⁸is chosen from H and (C₁-C₆) alkyl, or, azetidinyl, piperidinyl,oxadiazolyl, cyclopropyl, cyclobutyl, cyclopentyl, substitutedazetidinyl, substituted piperidinyl, substituted oxadiazolyl,substituted cyclopropyl, substituted cyclobutyl, substitutedcyclopentyl; R⁹ is chosen from hydrogen, alkyl, and substituted alkyl;and y is zero or an integer selected from 1, 2, and
 3. 2. A compoundaccording to claim 1, wherein R⁹ is hydrogen and R¹ is chosen from aryl,substituted aryl, azetidinyl, piperidinyl, piperazinyl, pyridinyl,pyrimidinyl, morpholinyl, thiomorpholinyl, pyrrolyl, pyrazolyl, indolyl,isoquinolinyl, azepinyl, diazepinyl, cyclobutyl, and substitutedazetidinyl, substituted piperidinyl, substituted piperazinyl,substituted pyridinyl, substituted pyrimidinyl, substituted morpholinyl,substituted thiomorpholinyl, substituted pyrrolyl, substitutedpyrazolyl, substituted indolyl, substituted isoquinolinyl, substitutedazepinyl, substituted diazepinyl, substituted cyclobutyl.
 3. A compoundaccording to claim 2, wherein R¹ is chosen from pyridinyl, pyrazolyl,pyrimidinyl, isoquinolinyl, azetidinyl, piperidinyl, piperizinyl,pyrrolidinyl, morpholinyl, azepanyl, diazepanyl, and phenyl optionallysubstituted with hydroxy, halogen, carboxamido, alkyl, carboxy, sulfone,alkoxy, and cyano.
 4. A compound according to claim 1, wherein R¹ ischosen from aryl, substituted aryl, (C₁-C₆) alkyl, and substituted(C₁-C₆) alkyl.
 5. A compound according to claim 1, wherein R¹ is —V—R⁷.6. A compound according to claim 5, wherein V is —C(═O)O— or —C(═O)NR⁸—,and R⁷ is chosen from —CH₂CN, (C₁-C₆) alkyl, and H, or VR⁷ is


7. A compound according to claim 5, wherein V is chosen from —O— and—NR⁸—; R⁷ is (C₁-C₆) alkyl; and R⁸ is H.
 8. A compound according toclaim 1, wherein R⁹ is hydrogen, y is zero and R⁴ is a residue selectedfrom a monocycle, a bicycle, a substituted monocycle, and a substitutedbicycle said residue containing at least one oxygen atom.
 9. A compoundaccording to claim 8, wherein R⁴ is chosen from pyranyl, benzopyranyl,furanyl, substituted pyranyl, substituted benzopyranyl, substitutedfuranyl, a hydroxyl-substituted cycloalkyl, an alkoxy-substituted aryl,and a hydroxyl-substituted aryl.
 10. A compound according to claim 9,wherein R⁴ is chosen from

wherein R⁶ is hydrogen or fluorine, and the carbon marked with anasterisk is of the R absolute configuration.
 11. A compound according toclaim 1, wherein R⁹ is hydrogen, y is 1-3, R² and R³ are hydrogen in alloccurrences, and R⁴ is alkoxy or OH of formula Ib or IIb:

wherein R⁵ is hydrogen or (C₁-C₆) alkyl.
 12. A compound according toclaim 1, wherein R⁹ is hydrogen, Q is CX, and y is zero of formula Ic orIIc:


13. A compound according to claim 12, wherein R¹ is chosen from halogen,substituted (C₁-C₆) alkyl, azetidinyl, piperidinyl, piperazinyl,pyridinyl, pyrimidinyl, morpholinyl, thiomorpholinyl, pyrrolyl,pyrazolyl, indolyl, isoquinolinyl, azepinyl, diazepinyl, cyclobutyl,substituted azetidinyl, substituted piperidinyl, substitutedpiperazinyl, substituted pyridinyl, substituted pyrimidinyl, substitutedmorpholinyl, substituted thiomorpholinyl, substituted pyrrolyl,substituted pyrazolyl, substituted indolyl, substituted isoquinolinyl,substituted azepinyl, substituted diazepinyl, substituted cyclobutyl,aryl, substituted aryl, and —V—R⁷.
 14. A compound according to claim 13,wherein V is —NR⁸—, —C(═O)O—, —C(═O)NR⁸, or O; R⁷ is H, (C₁-C₆) alkyl,substituted (C₁-C₆) alkyl, pyridinyl, piperidinyl, furanyl, oxadiazolyl,azetidinyl, cyclopropyl, cyclobutyl, cyclopentyl, substituted pyridinyl,substituted piperidinyl, substituted furanyl, substituted oxadiazolyl,substituted azetidinyl, substituted cyclopropyl, substituted cyclobutyl,substituted cyclopentyl, or aryl; and R⁸ is hydrogen or (C₁-C₆) alkyl,or azetidinyl, piperidinyl, oxadiazolyl, cyclopropyl, cyclobutyl,cyclopentyl, substituted azetidinyl, substituted piperidinyl,substituted oxadiazolyl, substituted cyclopropyl, substitutedcyclobutyl, substituted cyclopentyl.
 15. A compound according to claim12, wherein R⁴ is chosen from tetrahydrofuranyl, pyranyl, benzopyranyl,hydroxytetralinyl, oxepanyl, hydroxycyclohexyl, and their halogenatedcongeners; R¹ is chosen from halogen, (C₂-C₆) alkyl, substituted (C₁-C₆)alkyl, azetidinyl, phenyl, piperidinyl, piperizinyl, pyrrolidinyl,morpholinyl, azepanyl, diazepanyl, pyridinyl, pyrimidinyl, and pyrazolyloptionally substituted with hydroxy, halogen, carboxamido, alkyl,carboxy, sulfonyl, alkoxy, and cyano; and X is chosen from halogen,cyano, substituted alkoxy, and hydrogen.
 16. A compound according toclaim 15 selected from the group consisting of:


17. A compound according to claim 14 selected from the group consistingof:


18. A compound according to claim 1, wherein R⁹ is hydrogen, Q is CX, yis zero, and R⁴ is hydroxytetralinyl and hydroxycyclohexyl of formula Idor IId:

wherein R⁶ is H or halogen, and R¹ is chosen from halogen, azetidinyl,piperidinyl, piperizinyl, pyrrolidinyl, phenyl, morpholinyl, azepanyl,diazepanyl, pyridinyl pyrimidinyl, and pyrazolyl optionally substitutedwith hydroxy, halogen, carboxamido, alkyl, carboxy, sulfonyl, alkoxy,and cyano; and X is halogen, cyano, substituted alkoxy, and hydrogen.19. A compound according to claim 18 selected from


20. A compound according to claim 1, wherein Q is CX, y is zero, and R⁹is alkyl or substituted alkyl of formula Ie:


21. A compound according to claim 20, wherein R⁴ is chosen fromtetrahydrofuran, benzopyran, hydroxytetralin, oxepane,hydroxycyclohexane, and their halogenated congeners.
 22. A compoundaccording to claim 21, wherein R¹ is chosen from halogen, heteroaryl,heterocyclyl, substituted (C₁-C₆) alkyl, aryl, substituted aryl, cyano,carboxy, carboalkoxy, carboxamido, and amidino; and X is chosen fromhalogen, cyano, hydrogen, or alkoxy.
 23. A compound according to claim22 selected from the group consisting of:


24. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier and a therapeutically effective amount of a compoundaccording to claim 1.